The GeneSwitch system (GS) is a genetic reagent for pharmacologically inducible gene expression in Drosophila using the Gal4-UAS binary expression system. We have identified that the pan-glial repo-GS-Gal4 line uniquely causes significant baseline sleep reduction during both day and night in the presence of the inducing agent RU-486 even in the absence of effector transgene expression. Careful experimental design to permit detection of small sleep changes that may be obscured by the large sleep reduction in the presence of RU-486 is required when using the repo-GS-Gal4 line for behavioral studies.
","acknowledgements":"Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study.
","authors":[{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute",""],"credit":["dataCuration","formalAnalysis","methodology","writing_originalDraft","writing_reviewEditing","investigation"],"email":"sek155@rutgers.edu","firstName":"Seanna E","lastName":"Kelly","submittingAuthor":true,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0005-2726-3665"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["dataCuration","formalAnalysis","methodology","investigation","visualization","writing_originalDraft","writing_reviewEditing"],"email":"kf517@waksman.rutgers.edu","firstName":"Keisuke","lastName":"Fukumura","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0001-0126-0344"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["formalAnalysis","investigation"],"email":"dvm46@scarletmail.rutgers.edu","firstName":"Daridh","lastName":"Mao","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0007-6126-0278"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["investigation"],"email":"vinithra2021@gmail.com","firstName":"Vinithra","lastName":"Kathirvel","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":null},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["investigation"],"email":"sm2227@rwjms.rutgers.edu","firstName":"Shorbon","lastName":"Mowla","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0001-6559-0967"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["fundingAcquisition","writing_reviewEditing","supervision","project","conceptualization","resources"],"email":"annika.barber@waksman.rutgers.edu","firstName":"Annika F ","lastName":"Barber","submittingAuthor":false,"correspondingAuthor":true,"equalContribution":false,"WBId":null,"orcid":"0000-0002-5058-939X"}],"awards":[],"conflictsOfInterest":"The authors declare that there are no conflicts of interest present.
","dataTable":{"url":null},"extendedData":[],"funding":"We acknowledge National Institutes of Health R21 NS135530 (A.F.B.) and New Jersey Commission on Brain Injury Research CBIR24FEL012 (S.E.K.) for funding support.
","image":{"url":"https://portal.micropublication.org/uploads/c19ccc82cdf8d7fadf4f397d7d911cfa.png"},"imageCaption":"(A) Quantification of 24-h sleep time over 7 days for ;repo-GeneSwitch-Gal4/+; flies fed 0 (vehicle control), 50, 100, 200, or 465 µM concentration of RU-486 food. Asterisks denote significant difference in sleep time from vehicle control. (B) Relative gfp mRNA expression normalized to a-tub in heads of ;repo-GeneSwitch-Gal4/UAS-nls.GFP; flies fed 0 (vehicle control), 25, 50, 100, 200, or 465 µM RU-486. Asterisks denote significant difference GFP expression from vehicle control, there were no statistically significant differences between drug-fed groups. (C) As in (A) for ;;nSyb-GeneSwitch-Gal4/+ flies. Asterisks denote significant difference in sleep time from vehicle control. (D) As in (B) for ;UAS-nls.GFP/+;nSyb-GeneSwitch-Gal4/+ flies. Asterisks denote significant differences for the indicated comparisons.
","imageTitle":"repo-GeneSwitch-Gal4 substantially reduces baseline sleep at all concentrations of RU-486 significantly alters baseline sleep
","methods":"Fly Husbandry
Fly stocks used were repo-GS-Gal4 (BDSC # 95307), nSyb-GS-Gal4 (Bedont et al., 2021), UAS-nls.GFP (BDSC # 4776), and Iso31 (BDSC #5905). Geneswitch-Gal4 lines were back-crossed to Iso31 for 6 generations to normalize genetic background. All stocks were maintained on Bloomington Drosophila Stock Center cornmeal food in a 12:12 light-dark cycle at 25° C.
Drosophila Activity Monitor (DAM) Assay
repo-GS-Gal4 and nSyb-GS-Gal4 virgins were collected and crossed to Iso31 males. Male ;repo-GS-gal4/+; and ;;nSyb-GS-Gal4/+ progeny were collected 2-3 days post-eclosion and were placed in vials containing RU-486 DAM food (5% sucrose and 2% agar) or DAM food with 80% ethanol vehicle. Flies were placed on RU-486 food for 48 hours prior to being loaded into the DAM assay. Flies were placed in individual DAM tubes containing RU-486 or vehicle DAM food in one end and a cotton plug in the other. Flies were monitored in the assay for 7 days in a 12:12 light-dark cycle at 25° C. Sleep analysis was conducted using PHASE (Persons et al., 2022) and one-way ANOVA with Tukey’s post hoc test was used to determine statistical significance.
RU-486 Stock Dilutions
RU-486 is from Tokyo Chemical Industry (TCI), catalogue # TCI-M1732-1G. RU-486 was resuspended in molecular grade ethanol (80%) for a stock solution of 20 mg/mL. Working solution was further diluted to 20 mg/mL in 80% ethanol. All solutions were stored at -20° C.
Fly head qPCR
Total RNA was extracted from 50 heads per biological replicate, collected from 10-11-day old adult males maintained on RU-486 or vehicle diet for 7 days prior to sample collection. Tissues were homogenized in TRIzol™ Reagent (Invitrogen, Carlsbad, CA) and purified using the PureLink RNA Micro Kit (Invitrogen) with on-column PureLink DNase treatment according to the manufacturer's protocol. 100 ng of RNA was reverse transcribed using SuperScript IV Reverse Transcriptase (Invitrogen). Quantitative RT-PCR was performed on a CFX Opus 384 Real-Time PCR System (Bio-Rad, Hercules, CA) using iTaq™ Universal SYBR® Green Supermix (Bio-Rad). Cycling conditions were 95° C for 3 m, followed by 40 cycles of 95° C for 10 s and 60° C for 30 s, followed by melt curve analysis. Each sample was run in 3 technical replicates across 2–3 biological replicates. Relative expression levels were calculated by the DDCt method and normalized to a-tubulin. Sequences of primers are as follows: a-tubulin-Fw 5'-CGTCTGGACCACAAGTTCGA-3', a-tubulin-Rv 5'-CCTCCATACCCTCACCAACGT-3', gfp-Fw 5'-ATTGGCGATGGCCCTGTCCT-3', gfp-Rv 5'-GTTCATCCATGCCATGTGTAATCC-3'.
","reagents":"","patternDescription":"Inducible gene expression tools in Drosophila are widely used within the fly community for temporal control of gene expression. One such system is GeneSwitch (GS), an inducible Gal4 system in which Gal4 is inactive under baseline conditions and activated upon feeding the progesterone antagonist RU-486 (Osterwalder et al., 2001). Within the fly sleep field, GS-Gal4s have been used to induce adult-specific changes in gene expression followed by evaluation of changes in sleep amount and sleep architecture to determine ongoing adult roles for genes regulating sleep (Artiushin et al., 2018; Axelrod et al., 2023; Cho et al., 2026; Han et al., 2024; Joiner et al., 2006; Kuo et al., 2010; F. Li et al., 2023; Q. Li & Stavropoulos, 2016; Y. Li et al., 2023, 2024; Ly et al., 2020; Pyfrom et al., 2025; Tabuchi et al., 2015; Wu et al., 2009). Gal4-GS expression or RU-486 feeding alone do not affect sleep behavior in adult flies, although RU-486 exhibits developmental toxicity (Afonso et al., 2015; Q. Li & Stavropoulos, 2016; Wu et al., 2009). Several studies have noted that repo-GS-Gal4 flies have reduced sleep compared to UAS control lines when fed RU-486 (F. Li et al., 2023; Y. Li et al., 2023; Pyfrom et al., 2025). Similar sleep reductions have also been noted using the and MB-GS-Gal4 (mushroom body) driver, but not the nSyb-GS-Gal4 (pan-neuronal) and da-GS-Gal4 (ubiquitous) drivers (Artiushin et al., 2018; Cho et al., 2026; Y. Li et al., 2023; Ly et al., 2020; Tabuchi et al., 2015; Wu et al., 2009). Concentrations of RU-486 used for sleep studies vary widely, from 100 µM to 5 mM, yet sleep deficits occurred in repo-GS-Gal4 and MB-GS-Gal4 flies across concentrations and time spent on RU-486 (Artiushin et al., 2018; Axelrod et al., 2023; Joiner et al., 2006; Pyfrom et al., 2025; Wu et al., 2009).
Just two prior studies have compared sleep metrics of repo-GS-Gal4 in the presence and absence of RU-486 (Cho et al., 2026; Pyfrom et al., 2025). Cho et al. (2026) find ~100 minutes of sleep loss per day for repo-GS-Gal4 flies on 0.5 mM RU-486, though the profile of sleep loss over time on RU-486 is not shown. Pyfrom et al. (2025) show average 24 h sleep profiles identifying substantial day- and nighttime sleep loss for repo-GS-Gal4 in the presence of 100 µM RU-486 compared to vehicle control. We sought to identify an RU-486 concentration at which repo-GS-Gal4 would have minimal sleep loss, while still inducing gene expression. We found that repo-GS-Gal4 has significantly and substantially reduced sleep at all RU-486 concentrations, a property unique to this GS line. The strong reduction in baseline sleep in repo-GS-Gal4 flies in the presence of RU-486 raises concerns around the utility of the system for genetic screens of sleep modulation.
We characterized baseline sleep behavior and transgene expression levels using the pan-glial repo-GS-Gal4 and pan-neuronal nSyb-GS-Gal4 lines at multiple RU-486 concentrations. Using either repo-GS-Gal4 or nSyb-GS-Gal4, we measured baseline sleep with the Drosophila activity monitoring (DAM) assay and evaluated UAS-transgene expression levels by q-PCR at varying RU-486 concentrations. Both GS lines were backcrossed to our w1118 isogenic line (Iso31) (Ryder et al., 2004) for at least 6 generations to control for genetic background. Flies were placed on either ethanol vehicle control, 25, 50, 100, 200, or 465 µM concentrations of RU-486 mixed into agar/sucrose food for 24-hours prior to sleep experiments or into Bloomington fly diet for qPCR analysis one week prior to assay.
Upon examining baseline sleep over 7 days, we found that RU-486 significantly and substantially reduced sleep in repo-GS-Gal4 flies. 24-hour sleep time was significantly reduced at all concentrations of RU-486 for repo-GS-Gal4 flies compared to vehicle control (Figure 1A). Baseline 24-h sleep of repo-GS-Gal4 flies on vehicle control had a 7-day average of 1140 ± 9 minutes and average sleep time decreased dose-dependently with RU-486 concentration: 300 ± 23 minutes loss at 50 and 100 µM, 400 ± 23 minutes loss at 200 µM, and 550 ± 14 minutes loss at 465 µM. As in other studies, we found that RU-486-induced sleep loss in repo-GS-Gal4 flies was larger in the day (zeitgeber time 0-12) than at night (zeitgeber time 12-24), however sleep loss was significant during both periods. During the day, flies lost ~300 minutes of sleep at all RU-486 concentrations, while at night sleep loss was more concentration dependent, with ~300 minutes of sleep loss at 200 µM RU-486 but only ~150 minutes at 50 or 200 µM RU-486. We noted that the effect of RU-486 feeding on sleep loss is ameliorated over time in the assay. Despite this amelioration, by day 7 in the assay, flies fed 200-465 µM RU-486 still lost significantly more sleep than vehicle controls.
These data suggest that sleep quantity in repo-GS-Gal4 flies is highly sensitive to RU-486 feeding even in the absence of transgene expression. qPCR analysis of repo-GS-Gal4-driven UAS-nls.GFP expression after 24 h of RU-486 feeding revealed that the repo-GS system is highly sensitive to RU-486 induction. All RU-486 concentrations tested resulted in maximal GFP in fly heads, with no significant difference in GFP expression from 25-465 µM RU-486 (Figure 1B). Our qPCR findings indicate that the repo-GS system is highly sensitive to even very low levels of RU-486 feeding for just 24 h.
The large sleep reducing effects of RU-486 do not extend across all GS-Gal4 lines. nSyb-GS flies still displayed a significant sleep reduction in response to RU-486 feeding on the first four days in the assay (Figure 1C). However, the absolute amount of sleep lost in nSyb-GS flies is only ~200 minutes/day over each of the first four days at all RU-486 concentrations tested Baseline 24-hr sleep of nSyb-GS flies on the vehicle control averages 1097 minutes over 7 days in the assay and does not show dose dependent decreases with RU-486 concentration: 100-minute sleep loss (± 9-12 minutes) for 50, 100, and 200 µM. Although RU-486 induces significant sleep loss in nSyb-GS flies over the first few days in the DAM assay, the absolute amount of sleep lost is never more than 200 minutes per day, compared to the loss of 500-800 minutes per day in repo-GS-Gal4 flies. Sleep loss in nSyb-GS flies is also time-of-day dependent, with more sleep loss at night than during the day at all RU-486 concentrations tested. UAS-nls.GFP induction in heads by nSyb-GS-Gal4 is dose dependent with at least 100 µM RU-486 required for a significant induction compared to vehicle control (Figure 1D).
Our findings highlight that repo-GS-Gal4 is uniquely sensitive to RU-486-induced sleep reduction in the absence of transgene induction. Previous studies found that repo-GS-Gal4 is leaky, with repo-GS-driven expression of green fluorescent protein (GFP) detected in some glial subtypes in the absence of RU-486 feeding (Artiushin et al., 2018). This leakiness is unlikely to underlie observed sleep reduction in the presence of repo-GS-Gal4 with RU-486, as there is no UAS-transgene available to express. The effect of repo-GS-Gal4 could be positional, as it is an insertion of a fragment of the glial-specific reversed polarity transcription factor gene inserted into the AttP40 docking site on chromosome II while nSyb-GS-Gal4 is inserted into the AttP2 site on Chromosome III. Regardless of the underlying cause, using repo-GS-Gal4 to assess sleep changes requires careful use of both genetic and pharmacological controls to ensure that sleep changes can be detected at the concentration of RU-486 used. In some cases, the large sleep reductions induced by RU-486 feeding alone may obscure small differences in sleep induced by transgene expression. Alternatives to the GS system should be used to validate sleep findings using the repo-GS system, such as the temperature sensitive Gal80 or Q systems. Previous studies in Drosophila have successfully used these inducible gene expression approaches to analyze sleep behavior (Q. Li & Stavropoulos, 2016; Y. Li et al., 2023). Other studies have used the GS system to measure debris clearance, axotomy, short- and long-term memory, and synaptic remodeling (Chang et al., 2025; Dissel et al., 2015; Jacobs & Sehgal, 2020; Szabó et al., 2023; Vincze et al., 2026). Given our observations of the effect of repo-GS-Gal4 on sleep, careful use of controls is warranted for any phenotype, as short sleep could indirectly affect other outcomes.
","references":[{"reference":"Afonso DJS, Liu D, Machado DR, Pan H, Jepson JEC, Rogulja D, Koh K. 2015. TARANIS Functions with Cyclin A and Cdk1 in a Novel Arousal Center to Control Sleep in Drosophila. Current Biology. 25: 1717.","pubmedId":"","doi":"10.1016/j.cub.2015.05.037"},{"reference":"Artiushin G, Zhang SL, Tricoire H, Sehgal A. 2018. Endocytosis at the Drosophila blood–brain barrier as a function for sleep. eLife. 7: e43326.","pubmedId":"","doi":"10.7554/eLife.43326"},{"reference":"Axelrod S, Li X, Sun Y, Lincoln S, Terceros A, O Neil J, et al., Young MW. 2023. The Drosophila blood–brain barrier regulates sleep via Moody G protein-coupled receptor signaling. Proceedings of the National Academy of Sciences. 120: e2309331120.","pubmedId":"","doi":"10.1073/pnas.2309331120"},{"reference":"Bedont JL, Toda H, Shi M, Park CH, Quake C, Stein C, Kolesnik A, Sehgal A. 2021. Short and long sleeping mutants reveal links between sleep and macroautophagy. eLife. 10: e64140.","pubmedId":"","doi":"10.7554/eLife.64140"},{"reference":"Chang YC, Peng YJ, Lee JY, Wen A, Chang KT. 2025. Peripheral glia and neurons jointly regulate activity-induced synaptic remodeling at the Drosophila neuromuscular junction. eLife. 14: RP104126.","pubmedId":"","doi":"10.7554/eLife.104126.3"},{"reference":"Cho B, Youngstrom DE, Killiany S, Guevara C, Randolph CE, Beveridge CH, et al., Sehgal A. 2026. Sleep-dependent clearance of brain lipids by peripheral blood cells. Nature. 651: 720.","pubmedId":"","doi":"10.1038/s41586-025-10050-w"},{"reference":"Dissel S, Angadi V, Kirszenblat L, Suzuki Y, Donlea J, Klose M, et al., Shaw PJ. 2015. Sleep Restores Behavioral Plasticity to Drosophila Mutants. Current Biology. 25: 1270.","pubmedId":"","doi":"10.1016/j.cub.2015.03.027"},{"reference":"Han E, Lee SS, Park KH, Blum ID, Liu Q, Mehta A, et al., Wu MN. 2024. Tob Regulates the Timing of Sleep Onset at Night in Drosophila. The Journal of Neuroscience: e0389232024.","pubmedId":"","doi":"10.1523/JNEUROSCI.0389-23.2024"},{"reference":"Jacobs JA, Sehgal A. 2020. Anandamide Metabolites Protect against Seizures through the TRP Channel Water Witch in Drosophila melanogaster. Cell Reports. 31: 107710.","pubmedId":"","doi":"10.1016/j.celrep.2020.107710"},{"reference":"Joiner WJ, Crocker A, White BH, Sehgal A. 2006. Sleep in Drosophila is regulated by adult mushroom bodies. Nature. 441: 757.","pubmedId":"","doi":"10.1038/nature04811"},{"reference":"Li F, Artiushin G, Sehgal A. 2023. Modulation of sleep by trafficking of lipids through the Drosophila blood-brain barrier. eLife. 12: e86336.","pubmedId":"","doi":"10.7554/eLife.86336"},{"reference":"Li Q, Stavropoulos N. 2016. Evaluation of Ligand-Inducible Expression Systems for Conditional Neuronal Manipulations of Sleep in Drosophila. G3 Genes|Genomes|Genetics. 6: 3351.","pubmedId":"","doi":"10.1534/g3.116.034132"},{"reference":"Li Y, Chouhan NS, Zhang SL, Moore RS, Noya SB, Shon J, Yue Z, Sehgal A. 2024. Modulation of RNA processing genes during sleep-dependent memory. eLife. 12: RP89023.","pubmedId":"","doi":"10.7554/eLife.89023.4"},{"reference":"Li Y, Haynes P, Zhang SL, Yue Z, Sehgal A. 2023. Ecdysone acts through cortex glia to regulate sleep in Drosophila. eLife. 12: e81723.","pubmedId":"","doi":"10.7554/eLife.81723"},{"reference":"Li Y, Zhou Z, Zhang X, Tong H, Li P, Zhang ZC, et al., Han J. 2013. Drosophila Neuroligin 4 Regulates Sleep through Modulating GABA Transmission. The Journal of Neuroscience. 33: 15545.","pubmedId":"","doi":"10.1523/JNEUROSCI.0819-13.2013"},{"reference":"Ly S, Lee DA, Strus E, Prober DA, Naidoo N. 2020. Evolutionarily Conserved Regulation of Sleep by the Protein Translational Regulator PERK. Current Biology. 30: 1639.","pubmedId":"","doi":"10.1016/j.cub.2020.02.030"},{"reference":"Osterwalder T, Yoon KS, White BH, Keshishian H. 2001. A conditional tissue-specific transgene expression system using inducible GAL4. Proceedings of the National Academy of Sciences. 98: 12596.","pubmedId":"","doi":"10.1073/pnas.221303298"},{"reference":"Persons JL, Abhilash L, Lopatkin AJ, Roelofs A, Bell EV, Fernandez MP, Shafer OT. 2022. PHASE: An Open-Source Program for the Analysis of Drosophila Ph ase, A ctivity, and S leep Under E ntrainment. Journal of Biological Rhythms. 37: 455.","pubmedId":"","doi":"10.1177/07487304221093114"},{"reference":"Pyfrom ES, Beveridge C, Haynes PR, Kanigicherla VA, Randolph CE, Carvalho Costa P, et al., Sehgal A. 2025. Neutral lipid processing in glia is sexually dimorphic and promotes sleep through diacylglycerol catabolism.","pubmedId":"","doi":"10.1101/2025.09.10.674993"},{"reference":"Ryder E, Blows F, Ashburner M, Bautista Llacer R, Coulson D, Drummond J, et al., Russell S. 2004. The DrosDel Collection. Genetics. 167: 797.","pubmedId":"","doi":"10.1534/genetics.104.026658"},{"reference":"Szabo, Vincze V, Chhatre AS, Jipa A, Bognar S, Varga KE, et al., Juhasz G. 2023. LC3-associated phagocytosis promotes glial degradation of axon debris after injury in Drosophila models. Nature Communications. 14: 3077.","pubmedId":"","doi":"10.1038/s41467-023-38755-4"},{"reference":"Tabuchi M, Lone SR, Liu S, Liu Q, Zhang J, Spira AP, Wu MN. 2015. Sleep Interacts with Aβ to Modulate Intrinsic Neuronal Excitability. Current Biology. 25: 702.","pubmedId":"","doi":"10.1016/j.cub.2015.01.016"},{"reference":"Vincze V, Eskudt Z, Feher Juhasz E, Chhatre AS, Jipa A, Galambos AR, et al., Szabo. 2026. Selective autophagy fine-tunes Stat92E activity by degrading Su(var)2-10/PIAS in Drosophila glia. Life Science Alliance. 9: e202503375.","pubmedId":"","doi":"10.26508/lsa.202503375"},{"reference":"Wu MN, Ho K, Crocker A, Yue Z, Koh K, Sehgal A. 2009. The Effects of Caffeine on Sleep in Drosophila Require PKA Activity, But Not the Adenosine Receptor. The Journal of Neuroscience. 29: 11029.","pubmedId":"","doi":"10.1523/JNEUROSCI.1653-09.2009"}],"title":"Drosophila pan-glial inducible Gal4 line alters baseline sleep
","reviews":[{"reviewer":{"displayName":"Joseph Bedont"},"openAcknowledgement":false,"status":{"submitted":true}}],"curatorReviews":[{"curator":{"displayName":"FlyBase Curators"},"openAcknowledgement":false,"submitted":null}]},{"id":"792efd15-9062-4543-b906-47ed0f16321b","decision":"accept","abstract":"The GeneSwitch system (GS) is a genetic reagent for pharmacologically inducible gene expression in Drosophila using the Gal4-UAS binary expression system. We have identified that the pan-glial repo-GS-Gal4 line uniquely causes significant baseline sleep reduction during both day and night in the presence of the inducing agent RU-486 even in the absence of effector transgene expression. Careful experimental design to permit detection of small sleep changes that may be obscured by the large sleep reduction in the presence of RU-486 is required when using the repo-GS-Gal4 line for behavioral studies.
","acknowledgements":"Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study.
","authors":[{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute",""],"credit":["dataCuration","formalAnalysis","methodology","writing_originalDraft","writing_reviewEditing","investigation"],"email":"sek155@rutgers.edu","firstName":"Seanna E","lastName":"Kelly","submittingAuthor":true,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0005-2726-3665"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["dataCuration","formalAnalysis","methodology","investigation","visualization","writing_originalDraft","writing_reviewEditing"],"email":"kf517@waksman.rutgers.edu","firstName":"Keisuke","lastName":"Fukumura","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0001-0126-0344"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["formalAnalysis","investigation"],"email":"dvm46@scarletmail.rutgers.edu","firstName":"Daridh","lastName":"Mao","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0007-6126-0278"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["investigation"],"email":"vinithra2021@gmail.com","firstName":"Vinithra","lastName":"Kathirvel","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":null},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["investigation"],"email":"sm2227@rwjms.rutgers.edu","firstName":"Shorbon","lastName":"Mowla","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0001-6559-0967"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["fundingAcquisition","writing_reviewEditing","supervision","project","conceptualization","resources"],"email":"annika.barber@waksman.rutgers.edu","firstName":"Annika F ","lastName":"Barber","submittingAuthor":false,"correspondingAuthor":true,"equalContribution":false,"WBId":null,"orcid":"0000-0002-5058-939X"}],"awards":[],"conflictsOfInterest":"The authors declare that there are no conflicts of interest present.
","dataTable":{"url":null},"extendedData":[],"funding":"We acknowledge National Institutes of Health R21 NS135530 (A.F.B.) and New Jersey Commission on Brain Injury Research CBIR24FEL012 (S.E.K.) for funding support.
","image":{"url":"https://portal.micropublication.org/uploads/c19ccc82cdf8d7fadf4f397d7d911cfa.png"},"imageCaption":"(A) Quantification of 24-h sleep time over 7 days for ;repo-GeneSwitch-Gal4/+; flies fed 0 (vehicle control), 50, 100, 200, or 465 µM concentration of RU-486 food. Asterisks denote significant difference in sleep time from vehicle control. (B) Relative gfp mRNA expression normalized to a-tub in heads of ;repo-GeneSwitch-Gal4/UAS-nls.GFP; flies fed 0 (vehicle control), 25, 50, 100, 200, or 465 µM RU-486. Asterisks denote significant difference GFP expression from vehicle control, there were no statistically significant differences between drug-fed groups. (C) As in (A) for ;;nSyb-GeneSwitch-Gal4/+ flies. Asterisks denote significant difference in sleep time from vehicle control. (D) As in (B) for ;UAS-nls.GFP/+;nSyb-GeneSwitch-Gal4/+ flies. Asterisks denote significant differences for the indicated comparisons.
","imageTitle":"repo-GeneSwitch-Gal4 substantially reduces baseline sleep at all concentrations of RU-486
","methods":"Fly Husbandry
Fly stocks used were repo-GS-Gal4 (BDSC # 95307), nSyb-GS-Gal4 (Bedont et al., 2021), UAS-nls.GFP (BDSC # 4776), and Iso31 (BDSC #5905). Geneswitch-Gal4 lines were back-crossed to Iso31 for 6 generations to normalize genetic background. All stocks were maintained on Bloomington Drosophila Stock Center cornmeal food in a 12:12 light-dark cycle at 25° C.
Drosophila Activity Monitor (DAM) Assay
repo-GS-Gal4 and nSyb-GS-Gal4 virgins were collected and crossed to Iso31 males. Male ;repo-GS-gal4/+; and ;;nSyb-GS-Gal4/+ progeny were collected 2-3 days post-eclosion and were placed in vials containing RU-486 DAM food (5% sucrose and 2% agar) or DAM food with 80% ethanol vehicle. Flies were placed on RU-486 food for 48 hours prior to being loaded into the DAM assay. Flies were placed in individual DAM tubes containing RU-486 or vehicle DAM food in one end and a cotton plug in the other. Flies were monitored in the assay for 7 days in a 12:12 light-dark cycle at 25° C. Sleep analysis was conducted using PHASE (Persons et al., 2022) and one-way ANOVA with Tukey’s post hoc test was used to determine statistical significance.
RU-486 Stock Dilutions
RU-486 is from Tokyo Chemical Industry (TCI), catalogue # TCI-M1732-1G. RU-486 was resuspended in molecular grade ethanol (80%) for a stock solution of 20 mg/mL. Working solution was further diluted to 20 mg/mL in 80% ethanol. All solutions were stored at -20° C.
Fly head qPCR
Total RNA was extracted from 50 heads per biological replicate, collected from 10-11-day old adult males maintained on RU-486 or vehicle diet for 7 days prior to sample collection. Tissues were homogenized in TRIzol™ Reagent (Invitrogen, Carlsbad, CA) and purified using the PureLink RNA Micro Kit (Invitrogen) with on-column PureLink DNase treatment according to the manufacturer's protocol. 100 ng of RNA was reverse transcribed using SuperScript IV Reverse Transcriptase (Invitrogen). Quantitative RT-PCR was performed on a CFX Opus 384 Real-Time PCR System (Bio-Rad, Hercules, CA) using iTaq™ Universal SYBR® Green Supermix (Bio-Rad). Cycling conditions were 95° C for 3 m, followed by 40 cycles of 95° C for 10 s and 60° C for 30 s, followed by melt curve analysis. Each sample was run in 3 technical replicates across 2–3 biological replicates. Relative expression levels were calculated by the DDCt method and normalized to a-tubulin. Sequences of primers are as follows: a-tubulin-Fw 5'-CGTCTGGACCACAAGTTCGA-3', a-tubulin-Rv 5'-CCTCCATACCCTCACCAACGT-3', gfp-Fw 5'-ATTGGCGATGGCCCTGTCCT-3', gfp-Rv 5'-GTTCATCCATGCCATGTGTAATCC-3'.
","reagents":"","patternDescription":"Inducible gene expression tools in Drosophila are widely used within the fly community for temporal control of gene expression. One such system is GeneSwitch (GS), an inducible Gal4 system in which Gal4 is inactive under baseline conditions and activated upon feeding the progesterone antagonist RU-486 (Osterwalder et al., 2001) [citation]. Within the fly sleep field, GS-Gal4s have been used to induce adult-specific changes in gene expression followed by evaluation of changes in sleep amount and sleep architecture to determine ongoing adult roles for genes regulating sleep (Artiushin et al., 2018; Axelrod et al., 2023; Cho et al., 2026; Han et al., 2024; Joiner et al., 2006; Kuo et al., 2010; F. Li et al., 2023; Q. Li & Stavropoulos, 2016; Y. Li et al., 2023, 2024; Ly et al., 2020; Pyfrom et al., 2025; Tabuchi et al., 2015; Wu et al., 2009). Gal4-GS expression or RU-486 feeding alone do not affect sleep behavior in adult flies, although RU-486 exhibits developmental toxicity (Afonso et al., 2015; Q. Li & Stavropoulos, 2016; Wu et al., 2009). Several studies have noted that repo-GS-Gal4 flies have reduced sleep compared to UAS control lines when fed RU-486 (F. Li et al., 2023; Y. Li et al., 2023; Pyfrom et al., 2025). Similar sleep reductions have also been noted using the MB-GS-Gal4 (mushroom body) driver, but not the nSyb-GS-Gal4 (pan-neuronal) and da-GS-Gal4 (ubiquitous) drivers (Artiushin et al., 2018; Cho et al., 2026; Y. Li et al., 2023; Ly et al., 2020; Tabuchi et al., 2015; Wu et al., 2009). Concentrations of RU-486 used for sleep studies vary widely, from 100 µM to 5 mM, yet sleep deficits occurred in repo-GS-Gal4 and MB-GS-Gal4 flies across concentrations and time spent on RU-486 (Artiushin et al., 2018; Axelrod et al., 2023; Joiner et al., 2006; Pyfrom et al., 2025; Wu et al., 2009).
Just two prior studies have compared sleep metrics of repo-GS-Gal4 in the presence and absence of RU-486 (Cho et al., 2026; Pyfrom et al., 2025). Cho et al. (2026) find ~100 minutes of sleep loss per day for repo-GS-Gal4 flies on 0.5 mM RU-486, though the profile of sleep loss over time on RU-486 is not shown. Pyfrom et al. (2025) show average 24 h sleep profiles identifying substantial day- and nighttime sleep loss for repo-GS-Gal4 in the presence of 100 µM RU-486 compared to vehicle control. We sought to identify an RU-486 concentration at which repo-GS-Gal4 would have minimal sleep loss, while still inducing gene expression. We found that repo-GS-Gal4 has significantly and substantially reduced sleep at all RU-486 concentrations, a property unique to this GS line among those previously tested in the literature. The strong reduction in baseline sleep in repo-GS-Gal4 flies in the presence of RU-486 raises concerns around the utility of the system for genetic screens of sleep modulation.
We characterized baseline sleep behavior and transgene expression levels using the pan-glial repo-GS-Gal4 and pan-neuronal nSyb-GS-Gal4 lines at multiple RU-486 concentrations. Using either repo-GS-Gal4 or nSyb-GS-Gal4, we measured baseline sleep with the Drosophila activity monitoring (DAM) assay and evaluated UAS-transgene expression levels by q-PCR at varying RU-486 concentrations. Both GS lines were backcrossed to our w1118 isogenic line (Iso31) (Ryder et al., 2004) for at least 6 generations to control for genetic background. Flies were placed on either ethanol vehicle control, 25, 50, 100, 200, or 465 µM concentrations of RU-486 mixed into agar/sucrose food for 24-hours prior to sleep experiments or into Bloomington fly diet for qPCR analysis one week prior to assay.
Upon examining baseline sleep over 7 days, we found that RU-486 significantly and substantially reduced sleep in repo-GS-Gal4 flies. 24-hour sleep time was significantly reduced at all concentrations of RU-486 for repo-GS flies compared to vehicle control (Figure 1A). Baseline 24-h sleep of repo-GS flies on vehicle control had a 7-day average of 1140 ± 9 minutes and average sleep time decreased dose-dependently with RU-486 concentration: 300 ± 23 minutes loss at 50 and 100 µM, 400 ± 23 minutes loss at 200 µM, and 550 ± 14 minutes loss at 465 µM. As in other studies, we found that RU-486-induced sleep loss in repo-GS-Gal4 flies was larger in the day (zeitgeber time 0-12) than at night (zeitgeber time 12-24), however sleep loss was significant during both periods. During the day, flies lost ~300 minutes of sleep at all RU-486 concentrations, while at night sleep loss was more concentration dependent, with ~300 minutes of sleep loss at 200 µM RU-486 but only ~150 minutes at 50 or 100 µM RU-486. We noted that the effect of RU-486 feeding on sleep loss is ameliorated over time in the assay. Despite this amelioration, by day 7 in the assay, flies fed 200-465 µM RU-486 still lost significantly more sleep than vehicle controls.
These data suggest that sleep quantity in repo-GS-Gal4 flies is highly sensitive to RU-486 feeding even in the absence of transgene expression. qPCR analysis of repo-GS-Gal4-driven UAS-nls.GFP expression after 24 h of RU-486 feeding revealed that the repo-GS system is highly sensitive to RU-486 induction. All RU-486 concentrations tested resulted in maximal GFP in fly heads, with no significant difference in GFP expression from 25-465 µM RU-486 (Figure 1B). Our qPCR findings indicate that the repo-GS system is highly sensitive to even very low levels of RU-486 feeding for just 24 h.
The large sleep reducing effects of RU-486 do not extend across all GS-Gal4 lines. nSyb-GS flies still displayed a significant sleep reduction in response to RU-486 feeding on the first four days in the assay (Figure 1C). However, the absolute amount of sleep lost in nSyb-GS flies is only ~200 minutes/day over each of the first four days at all RU-486 concentrations tested. Baseline 24-hr sleep of nSyb-GS flies on the vehicle control averages 1097 minutes over 7 days in the assay and does not show dose dependent decreases with RU-486 concentration: 100-minute sleep loss (± 9-12 minutes) for 50, 100, and 200 µM. Although RU-486 induces significant sleep loss in nSyb-GS flies over the first few days in the DAM assay, the absolute amount of sleep lost is never more than 200 minutes per day, compared to the loss of 500-800 minutes per day in repo-GS-Gal4 flies. Sleep loss in nSyb-GS flies is also time-of-day dependent, with more sleep loss at night than during the day at all RU-486 concentrations tested. UAS-nls.GFP induction in heads by nSyb-GS-Gal4 is dose dependent with at least 100 µM RU-486 required for a significant induction compared to vehicle control (Figure 1D).
Our findings highlight that repo-GS-Gal4 is uniquely sensitive to RU-486-induced sleep reduction in the absence of transgene induction. Previous studies found that repo-GS-Gal4 is leaky, with repo-GS-driven expression of green fluorescent protein (GFP) detected in some glial subtypes in the absence of RU-486 feeding (Artiushin et al., 2018). This leakiness is unlikely to underlie observed sleep reduction in the presence of repo-GS-Gal4 with RU-486, as there is no UAS-transgene available to express. The effect of repo-GS-Gal4 could be positional, as it is an insertion of a fragment of the glial-specific reversed polarity transcription factor gene inserted into the AttP40 docking site on chromosome II while nSyb-GS-Gal4 is inserted into the AttP2 site on Chromosome III. Regardless of the underlying cause, using repo-GS to assess sleep changes requires careful use of both genetic and pharmacological controls to ensure that sleep changes can be detected at the concentration of RU-486 used. In some cases, the large sleep reductions induced by RU-486 feeding alone may obscure small differences in sleep induced by transgene expression. Alternatives to the GS system should be used to validate sleep findings using the repo-GS system, such as the temperature sensitive Gal80 or Q systems. Previous studies in Drosophila have successfully used these inducible gene expression approaches to analyze sleep behavior (Q. Li & Stavropoulos, 2016; Y. Li et al., 2023). Other studies have used the GS system to measure debris clearance, axotomy, short- and long-term memory, and synaptic remodeling (Chang et al., 2025; Dissel et al., 2015; Jacobs & Sehgal, 2020; Szabó et al., 2023; Vincze et al., 2026). Given our observations of the effect of repo-GS on sleep, careful use of controls is warranted for any phenotype, as short sleep could indirectly affect other outcomes.
","references":[{"reference":"Afonso DJS, Liu D, Machado DR, Pan H, Jepson JEC, Rogulja D, Koh K. 2015. TARANIS Functions with Cyclin A and Cdk1 in a Novel Arousal Center to Control Sleep in Drosophila. Current Biology. 25: 1717.","pubmedId":"","doi":"10.1016/j.cub.2015.05.037"},{"reference":"Artiushin G, Zhang SL, Tricoire H, Sehgal A. 2018. Endocytosis at the Drosophila blood–brain barrier as a function for sleep. eLife. 7: e43326.","pubmedId":"","doi":"10.7554/eLife.43326"},{"reference":"Axelrod S, Li X, Sun Y, Lincoln S, Terceros A, O Neil J, et al., Young MW. 2023. The Drosophila blood–brain barrier regulates sleep via Moody G protein-coupled receptor signaling. Proceedings of the National Academy of Sciences. 120: e2309331120.","pubmedId":"","doi":"10.1073/pnas.2309331120"},{"reference":"Bedont JL, Toda H, Shi M, Park CH, Quake C, Stein C, Kolesnik A, Sehgal A. 2021. Short and long sleeping mutants reveal links between sleep and macroautophagy. eLife. 10: e64140.","pubmedId":"","doi":"10.7554/eLife.64140"},{"reference":"Chang YC, Peng YJ, Lee JY, Wen A, Chang KT. 2025. Peripheral glia and neurons jointly regulate activity-induced synaptic remodeling at the Drosophila neuromuscular junction. eLife. 14: RP104126.","pubmedId":"","doi":"10.7554/eLife.104126.3"},{"reference":"Cho B, Youngstrom DE, Killiany S, Guevara C, Randolph CE, Beveridge CH, et al., Sehgal A. 2026. Sleep-dependent clearance of brain lipids by peripheral blood cells. Nature. 651: 720.","pubmedId":"","doi":"10.1038/s41586-025-10050-w"},{"reference":"Dissel S, Angadi V, Kirszenblat L, Suzuki Y, Donlea J, Klose M, et al., Shaw PJ. 2015. Sleep Restores Behavioral Plasticity to Drosophila Mutants. Current Biology. 25: 1270.","pubmedId":"","doi":"10.1016/j.cub.2015.03.027"},{"reference":"Han E, Lee SS, Park KH, Blum ID, Liu Q, Mehta A, et al., Wu MN. 2024. Tob Regulates the Timing of Sleep Onset at Night in Drosophila. The Journal of Neuroscience: e0389232024.","pubmedId":"","doi":"10.1523/JNEUROSCI.0389-23.2024"},{"reference":"Jacobs JA, Sehgal A. 2020. Anandamide Metabolites Protect against Seizures through the TRP Channel Water Witch in Drosophila melanogaster. Cell Reports. 31: 107710.","pubmedId":"","doi":"10.1016/j.celrep.2020.107710"},{"reference":"Joiner WJ, Crocker A, White BH, Sehgal A. 2006. Sleep in Drosophila is regulated by adult mushroom bodies. Nature. 441: 757.","pubmedId":"","doi":"10.1038/nature04811"},{"reference":"Li F, Artiushin G, Sehgal A. 2023. Modulation of sleep by trafficking of lipids through the Drosophila blood-brain barrier. eLife. 12: e86336.","pubmedId":"","doi":"10.7554/eLife.86336"},{"reference":"Li Q, Stavropoulos N. 2016. Evaluation of Ligand-Inducible Expression Systems for Conditional Neuronal Manipulations of Sleep in Drosophila. G3 Genes|Genomes|Genetics. 6: 3351.","pubmedId":"","doi":"10.1534/g3.116.034132"},{"reference":"Li Y, Chouhan NS, Zhang SL, Moore RS, Noya SB, Shon J, Yue Z, Sehgal A. 2024. Modulation of RNA processing genes during sleep-dependent memory. eLife. 12: RP89023.","pubmedId":"","doi":"10.7554/eLife.89023.4"},{"reference":"Li Y, Haynes P, Zhang SL, Yue Z, Sehgal A. 2023. Ecdysone acts through cortex glia to regulate sleep in Drosophila. eLife. 12: e81723.","pubmedId":"","doi":"10.7554/eLife.81723"},{"reference":"Li Y, Zhou Z, Zhang X, Tong H, Li P, Zhang ZC, et al., Han J. 2013. Drosophila Neuroligin 4 Regulates Sleep through Modulating GABA Transmission. The Journal of Neuroscience. 33: 15545.","pubmedId":"","doi":"10.1523/JNEUROSCI.0819-13.2013"},{"reference":"Ly S, Lee DA, Strus E, Prober DA, Naidoo N. 2020. Evolutionarily Conserved Regulation of Sleep by the Protein Translational Regulator PERK. Current Biology. 30: 1639.","pubmedId":"","doi":"10.1016/j.cub.2020.02.030"},{"reference":"Osterwalder T, Yoon KS, White BH, Keshishian H. 2001. A conditional tissue-specific transgene expression system using inducible GAL4. Proceedings of the National Academy of Sciences. 98: 12596.","pubmedId":"","doi":"10.1073/pnas.221303298"},{"reference":"Persons JL, Abhilash L, Lopatkin AJ, Roelofs A, Bell EV, Fernandez MP, Shafer OT. 2022. PHASE: An Open-Source Program for the Analysis of Drosophila Ph ase, A ctivity, and S leep Under E ntrainment. Journal of Biological Rhythms. 37: 455.","pubmedId":"","doi":"10.1177/07487304221093114"},{"reference":"Pyfrom ES, Beveridge C, Haynes PR, Kanigicherla VA, Randolph CE, Carvalho Costa P, et al., Sehgal A. 2025. Neutral lipid processing in glia is sexually dimorphic and promotes sleep through diacylglycerol catabolism.","pubmedId":"","doi":"10.1101/2025.09.10.674993"},{"reference":"Ryder E, Blows F, Ashburner M, Bautista Llacer R, Coulson D, Drummond J, et al., Russell S. 2004. The DrosDel Collection. Genetics. 167: 797.","pubmedId":"","doi":"10.1534/genetics.104.026658"},{"reference":"Szabo, Vincze V, Chhatre AS, Jipa A, Bognar S, Varga KE, et al., Juhasz G. 2023. LC3-associated phagocytosis promotes glial degradation of axon debris after injury in Drosophila models. Nature Communications. 14: 3077.","pubmedId":"","doi":"10.1038/s41467-023-38755-4"},{"reference":"Tabuchi M, Lone SR, Liu S, Liu Q, Zhang J, Spira AP, Wu MN. 2015. Sleep Interacts with Aβ to Modulate Intrinsic Neuronal Excitability. Current Biology. 25: 702.","pubmedId":"","doi":"10.1016/j.cub.2015.01.016"},{"reference":"Vincze V, Eskudt Z, Feher Juhasz E, Chhatre AS, Jipa A, Galambos AR, et al., Szabo. 2026. Selective autophagy fine-tunes Stat92E activity by degrading Su(var)2-10/PIAS in Drosophila glia. Life Science Alliance. 9: e202503375.","pubmedId":"","doi":"10.26508/lsa.202503375"},{"reference":"Wu MN, Ho K, Crocker A, Yue Z, Koh K, Sehgal A. 2009. The Effects of Caffeine on Sleep in Drosophila Require PKA Activity, But Not the Adenosine Receptor. The Journal of Neuroscience. 29: 11029.","pubmedId":"","doi":"10.1523/JNEUROSCI.1653-09.2009"}],"title":"Drosophila pan-glial inducible Gal4 line alters baseline sleep
","reviews":[],"curatorReviews":[{"curator":{"displayName":"FlyBase Curators"},"openAcknowledgement":false,"submitted":"1778740429575"}]},{"id":"b1ad62c0-3a2b-4871-9632-ad70d647de09","decision":"publish","abstract":"The GeneSwitch system (GS) is a genetic reagent for pharmacologically inducible gene expression in Drosophila using the Gal4-UAS binary expression system. We have identified that the pan-glial repo-GS-Gal4 line uniquely causes significant baseline sleep reduction during both day and night in the presence of the inducing agent RU-486 even in the absence of effector transgene expression. Careful experimental design to permit detection of small sleep changes that may be obscured by the large sleep reduction in the presence of RU-486 is required when using the repo-GS-Gal4 line for behavioral studies.
","acknowledgements":"Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study.
","authors":[{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute",""],"credit":["dataCuration","formalAnalysis","methodology","writing_originalDraft","writing_reviewEditing","investigation"],"email":"sek155@rutgers.edu","firstName":"Seanna E","lastName":"Kelly","submittingAuthor":true,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0005-2726-3665"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["dataCuration","formalAnalysis","methodology","investigation","visualization","writing_originalDraft","writing_reviewEditing"],"email":"kf517@waksman.rutgers.edu","firstName":"Keisuke","lastName":"Fukumura","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0001-0126-0344"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["formalAnalysis","investigation"],"email":"dvm46@scarletmail.rutgers.edu","firstName":"Daridh","lastName":"Mao","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0007-6126-0278"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["investigation"],"email":"vinithra2021@gmail.com","firstName":"Vinithra","lastName":"Kathirvel","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":null},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["investigation"],"email":"sm2227@rwjms.rutgers.edu","firstName":"Shorbon","lastName":"Mowla","submittingAuthor":false,"correspondingAuthor":false,"equalContribution":false,"WBId":null,"orcid":"0009-0001-6559-0967"},{"affiliations":["Rutgers, New Brunswick, NJ, US"],"departments":["Waksman Institute"],"credit":["fundingAcquisition","writing_reviewEditing","supervision","project","conceptualization","resources"],"email":"annika.barber@waksman.rutgers.edu","firstName":"Annika F ","lastName":"Barber","submittingAuthor":false,"correspondingAuthor":true,"equalContribution":false,"WBId":null,"orcid":"0000-0002-5058-939X"}],"awards":[],"conflictsOfInterest":"The authors declare that there are no conflicts of interest present.
","dataTable":{"url":null},"extendedData":[],"funding":"We acknowledge National Institutes of Health R21 NS135530 (A.F.B.) and New Jersey Commission on Brain Injury Research CBIR24FEL012 (S.E.K.) for funding support.
","image":{"url":"https://portal.micropublication.org/uploads/c19ccc82cdf8d7fadf4f397d7d911cfa.png"},"imageCaption":"(A) Quantification of 24-h sleep time over 7 days for ;repo-GeneSwitch-Gal4/+; flies fed 0 (vehicle control), 50, 100, 200, or 465 µM concentration of RU-486 food. Asterisks denote significant difference in sleep time from vehicle control. (B) Relative gfp mRNA expression normalized to a-tub in heads of ;repo-GeneSwitch-Gal4/UAS-nls.GFP; flies fed 0 (vehicle control), 25, 50, 100, 200, or 465 µM RU-486. Asterisks denote significant difference GFP expression from vehicle control, there were no statistically significant differences between drug-fed groups. (C) As in (A) for ;;nSyb-GeneSwitch-Gal4/+ flies. Asterisks denote significant difference in sleep time from vehicle control. (D) As in (B) for ;UAS-nls.GFP/+;nSyb-GeneSwitch-Gal4/+ flies. Asterisks denote significant differences for the indicated comparisons.
","imageTitle":"repo-GeneSwitch-Gal4 substantially reduces baseline sleep at all concentrations of RU-486
","methods":"Fly Husbandry
Fly stocks used were repo-GS-Gal4 (BDSC # 95307), nSyb-GS-Gal4 (Bedont et al., 2021), UAS-nls.GFP (BDSC # 4776), and Iso31 (BDSC #5905). Geneswitch-Gal4 lines were back-crossed to Iso31 for 6 generations to normalize genetic background. All stocks were maintained on Bloomington Drosophila Stock Center cornmeal food in a 12:12 light-dark cycle at 25° C.
Drosophila Activity Monitor (DAM) Assay
repo-GS-Gal4 and nSyb-GS-Gal4 virgins were collected and crossed to Iso31 males. Male ;repo-GS-gal4/+; and ;;nSyb-GS-Gal4/+ progeny were collected 2-3 days post-eclosion and were placed in vials containing RU-486 DAM food (5% sucrose and 2% agar) or DAM food with 80% ethanol vehicle. Flies were placed on RU-486 food for 48 hours prior to being loaded into the DAM assay. Flies were placed in individual DAM tubes containing RU-486 or vehicle DAM food in one end and a cotton plug in the other. Flies were monitored in the assay for 7 days in a 12:12 light-dark cycle at 25° C. Sleep analysis was conducted using PHASE (Persons et al., 2022) and one-way ANOVA with Tukey’s post hoc test was used to determine statistical significance.
RU-486 Stock Dilutions
RU-486 is from Tokyo Chemical Industry (TCI), catalogue # TCI-M1732-1G. RU-486 was resuspended in molecular grade ethanol (80%) for a stock solution of 20 mg/mL. Working solution was further diluted to 20 mg/mL in 80% ethanol. All solutions were stored at -20° C.
Fly head qPCR
Total RNA was extracted from 50 heads per biological replicate, collected from 10-11-day old adult males maintained on RU-486 or vehicle diet for 7 days prior to sample collection. Tissues were homogenized in TRIzol™ Reagent (Invitrogen, Carlsbad, CA) and purified using the PureLink RNA Micro Kit (Invitrogen) with on-column PureLink DNase treatment according to the manufacturer's protocol. 100 ng of RNA was reverse transcribed using SuperScript IV Reverse Transcriptase (Invitrogen). Quantitative RT-PCR was performed on a CFX Opus 384 Real-Time PCR System (Bio-Rad, Hercules, CA) using iTaq™ Universal SYBR® Green Supermix (Bio-Rad). Cycling conditions were 95° C for 3 m, followed by 40 cycles of 95° C for 10 s and 60° C for 30 s, followed by melt curve analysis. Each sample was run in 3 technical replicates across 2–3 biological replicates. Relative expression levels were calculated by the DDCt method and normalized to a-tubulin. Sequences of primers are as follows: a-tubulin-Fw 5'-CGTCTGGACCACAAGTTCGA-3', a-tubulin-Rv 5'-CCTCCATACCCTCACCAACGT-3', gfp-Fw 5'-ATTGGCGATGGCCCTGTCCT-3', gfp-Rv 5'-GTTCATCCATGCCATGTGTAATCC-3'.
","reagents":"","patternDescription":"Inducible gene expression tools in Drosophila are widely used within the fly community for temporal control of gene expression. One such system is GeneSwitch (GS), an inducible Gal4 system in which Gal4 is inactive under baseline conditions and activated upon feeding the progesterone antagonist RU-486 (Osterwalder et al., 2001) [citation]. Within the fly sleep field, GS-Gal4s have been used to induce adult-specific changes in gene expression followed by evaluation of changes in sleep amount and sleep architecture to determine ongoing adult roles for genes regulating sleep (Artiushin et al., 2018; Axelrod et al., 2023; Cho et al., 2026; Han et al., 2024; Joiner et al., 2006; Kuo et al., 2010; F. Li et al., 2023; Q. Li & Stavropoulos, 2016; Y. Li et al., 2023, 2024; Ly et al., 2020; Pyfrom et al., 2025; Tabuchi et al., 2015; Wu et al., 2009). Gal4-GS expression or RU-486 feeding alone do not affect sleep behavior in adult flies, although RU-486 exhibits developmental toxicity (Afonso et al., 2015; Q. Li & Stavropoulos, 2016; Wu et al., 2009). Several studies have noted that repo-GS-Gal4 flies have reduced sleep compared to UAS control lines when fed RU-486 (F. Li et al., 2023; Y. Li et al., 2023; Pyfrom et al., 2025). Similar sleep reductions have also been noted using the MB-GS-Gal4 (mushroom body) driver, but not the nSyb-GS-Gal4 (pan-neuronal) and da-GS-Gal4 (ubiquitous) drivers (Artiushin et al., 2018; Cho et al., 2026; Y. Li et al., 2023; Ly et al., 2020; Tabuchi et al., 2015; Wu et al., 2009). Concentrations of RU-486 used for sleep studies vary widely, from 100 µM to 5 mM, yet sleep deficits occurred in repo-GS-Gal4 and MB-GS-Gal4 flies across concentrations and time spent on RU-486 (Artiushin et al., 2018; Axelrod et al., 2023; Joiner et al., 2006; Pyfrom et al., 2025; Wu et al., 2009).
Just two prior studies have compared sleep metrics of repo-GS-Gal4 in the presence and absence of RU-486 (Cho et al., 2026; Pyfrom et al., 2025). Cho et al. (2026) find ~100 minutes of sleep loss per day for repo-GS-Gal4 flies on 0.5 mM RU-486, though the profile of sleep loss over time on RU-486 is not shown. Pyfrom et al. (2025) show average 24 h sleep profiles identifying substantial day- and nighttime sleep loss for repo-GS-Gal4 in the presence of 100 µM RU-486 compared to vehicle control. We sought to identify an RU-486 concentration at which repo-GS-Gal4 would have minimal sleep loss, while still inducing gene expression. We found that repo-GS-Gal4 has significantly and substantially reduced sleep at all RU-486 concentrations, a property unique to this GS line among those previously tested in the literature. The strong reduction in baseline sleep in repo-GS-Gal4 flies in the presence of RU-486 raises concerns around the utility of the system for genetic screens of sleep modulation.
We characterized baseline sleep behavior and transgene expression levels using the pan-glial repo-GS-Gal4 and pan-neuronal nSyb-GS-Gal4 lines at multiple RU-486 concentrations. Using either repo-GS-Gal4 or nSyb-GS-Gal4, we measured baseline sleep with the Drosophila activity monitoring (DAM) assay and evaluated UAS-transgene expression levels by q-PCR at varying RU-486 concentrations. Both GS lines were backcrossed to our w1118 isogenic line (Iso31) (Ryder et al., 2004) for at least 6 generations to control for genetic background. Flies were placed on either ethanol vehicle control, 25, 50, 100, 200, or 465 µM concentrations of RU-486 mixed into agar/sucrose food for 24-hours prior to sleep experiments or into Bloomington fly diet for qPCR analysis one week prior to assay.
Upon examining baseline sleep over 7 days, we found that RU-486 significantly and substantially reduced sleep in repo-GS-Gal4 flies. 24-hour sleep time was significantly reduced at all concentrations of RU-486 for repo-GS flies compared to vehicle control (Figure 1A). Baseline 24-h sleep of repo-GS flies on vehicle control had a 7-day average of 1140 ± 9 minutes and average sleep time decreased dose-dependently with RU-486 concentration: 300 ± 23 minutes loss at 50 and 100 µM, 400 ± 23 minutes loss at 200 µM, and 550 ± 14 minutes loss at 465 µM. As in other studies, we found that RU-486-induced sleep loss in repo-GS-Gal4 flies was larger in the day (zeitgeber time 0-12) than at night (zeitgeber time 12-24), however sleep loss was significant during both periods. During the day, flies lost ~300 minutes of sleep at all RU-486 concentrations, while at night sleep loss was more concentration dependent, with ~300 minutes of sleep loss at 200 µM RU-486 but only ~150 minutes at 50 or 100 µM RU-486. We noted that the effect of RU-486 feeding on sleep loss is ameliorated over time in the assay. Despite this amelioration, by day 7 in the assay, flies fed 200-465 µM RU-486 still lost significantly more sleep than vehicle controls.
These data suggest that sleep quantity in repo-GS-Gal4 flies is highly sensitive to RU-486 feeding even in the absence of transgene expression. qPCR analysis of repo-GS-Gal4-driven UAS-nls.GFP expression after 24 h of RU-486 feeding revealed that the repo-GS system is highly sensitive to RU-486 induction. All RU-486 concentrations tested resulted in maximal GFP in fly heads, with no significant difference in GFP expression from 25-465 µM RU-486 (Figure 1B). Our qPCR findings indicate that the repo-GS system is highly sensitive to even very low levels of RU-486 feeding for just 24 hours.
The large sleep reducing effects of RU-486 do not extend across all GS-Gal4 lines. nSyb-GS flies still displayed a significant sleep reduction in response to RU-486 feeding on the first four days in the assay (Figure 1C). However, the absolute amount of sleep lost in nSyb-GS flies is only ~200 minutes/day over each of the first four days at all RU-486 concentrations tested. Baseline 24-hr sleep of nSyb-GS flies on the vehicle control averages 1097 minutes over 7 days in the assay and does not show dose dependent decreases with RU-486 concentration: 100-minute sleep loss (± 9-12 minutes) for 50, 100, and 200 µM. Although RU-486 induces significant sleep loss in nSyb-GS flies over the first few days in the DAM assay, the absolute amount of sleep lost is never more than 200 minutes per day, compared to the loss of 500-800 minutes per day in repo-GS-Gal4 flies. Sleep loss in nSyb-GS flies is also time-of-day dependent, with more sleep loss at night than during the day at all RU-486 concentrations tested. UAS-nls.GFP induction in heads by nSyb-GS-Gal4 is dose dependent with at least 100 µM RU-486 required for a significant induction compared to vehicle control (Figure 1D).
Our findings highlight that repo-GS-Gal4 is uniquely sensitive to RU-486-induced sleep reduction in the absence of transgene induction. Previous studies found that repo-GS-Gal4 is leaky, with repo-GS-driven expression of green fluorescent protein (GFP) detected in some glial subtypes in the absence of RU-486 feeding (Artiushin et al., 2018). This leakiness is unlikely to underlie observed sleep reduction in the presence of repo-GS-Gal4 with RU-486, as there is no UAS-transgene available to express. The effect of repo-GS-Gal4 could be positional, as it is an insertion of a fragment of the glial-specific reversed polarity transcription factor gene inserted into the AttP40 docking site on chromosome II while nSyb-GS-Gal4 is inserted into the AttP2 site on Chromosome III. Regardless of the underlying cause, using repo-GS to assess sleep changes requires careful use of both genetic and pharmacological controls to ensure that sleep changes can be detected at the concentration of RU-486 used. In some cases, the large sleep reductions induced by RU-486 feeding alone may obscure small differences in sleep induced by transgene expression. Alternatives to the GS system should be used to validate sleep findings using the repo-GS system, such as the temperature sensitive Gal80 or Q systems. Previous studies in Drosophila have successfully used these inducible gene expression approaches to analyze sleep behavior (Q. Li & Stavropoulos, 2016; Y. Li et al., 2023). Other studies have used the GS system to measure debris clearance, axotomy, short- and long-term memory, and synaptic remodeling (Chang et al., 2025; Dissel et al., 2015; Jacobs & Sehgal, 2020; Szabó et al., 2023; Vincze et al., 2026). Given our observations of the effect of repo-GS on sleep, careful use of controls is warranted for any phenotype, as short sleep could indirectly affect other outcomes.
","references":[{"reference":"Afonso DJS, Liu D, Machado DR, Pan H, Jepson JEC, Rogulja D, Koh K. 2015. TARANIS Functions with Cyclin A and Cdk1 in a Novel Arousal Center to Control Sleep in Drosophila. Current Biology. 25: 1717.","pubmedId":"","doi":"10.1016/j.cub.2015.05.037"},{"reference":"Artiushin G, Zhang SL, Tricoire H, Sehgal A. 2018. Endocytosis at the Drosophila blood–brain barrier as a function for sleep. eLife. 7: e43326.","pubmedId":"","doi":"10.7554/eLife.43326"},{"reference":"Axelrod S, Li X, Sun Y, Lincoln S, Terceros A, O Neil J, et al., Young MW. 2023. The Drosophila blood–brain barrier regulates sleep via Moody G protein-coupled receptor signaling. Proceedings of the National Academy of Sciences. 120: e2309331120.","pubmedId":"","doi":"10.1073/pnas.2309331120"},{"reference":"Bedont JL, Toda H, Shi M, Park CH, Quake C, Stein C, Kolesnik A, Sehgal A. 2021. Short and long sleeping mutants reveal links between sleep and macroautophagy. eLife. 10: e64140.","pubmedId":"","doi":"10.7554/eLife.64140"},{"reference":"Chang YC, Peng YJ, Lee JY, Wen A, Chang KT. 2025. Peripheral glia and neurons jointly regulate activity-induced synaptic remodeling at the Drosophila neuromuscular junction. eLife. 14: RP104126.","pubmedId":"","doi":"10.7554/eLife.104126.3"},{"reference":"Cho B, Youngstrom DE, Killiany S, Guevara C, Randolph CE, Beveridge CH, et al., Sehgal A. 2026. Sleep-dependent clearance of brain lipids by peripheral blood cells. Nature. 651: 720.","pubmedId":"","doi":"10.1038/s41586-025-10050-w"},{"reference":"Dissel S, Angadi V, Kirszenblat L, Suzuki Y, Donlea J, Klose M, et al., Shaw PJ. 2015. Sleep Restores Behavioral Plasticity to Drosophila Mutants. Current Biology. 25: 1270.","pubmedId":"","doi":"10.1016/j.cub.2015.03.027"},{"reference":"Han E, Lee SS, Park KH, Blum ID, Liu Q, Mehta A, et al., Wu MN. 2024. Tob Regulates the Timing of Sleep Onset at Night in Drosophila. The Journal of Neuroscience: e0389232024.","pubmedId":"","doi":"10.1523/JNEUROSCI.0389-23.2024"},{"reference":"Jacobs JA, Sehgal A. 2020. Anandamide Metabolites Protect against Seizures through the TRP Channel Water Witch in Drosophila melanogaster. Cell Reports. 31: 107710.","pubmedId":"","doi":"10.1016/j.celrep.2020.107710"},{"reference":"Joiner WJ, Crocker A, White BH, Sehgal A. 2006. Sleep in Drosophila is regulated by adult mushroom bodies. Nature. 441: 757.","pubmedId":"","doi":"10.1038/nature04811"},{"reference":"Li F, Artiushin G, Sehgal A. 2023. Modulation of sleep by trafficking of lipids through the Drosophila blood-brain barrier. eLife. 12: e86336.","pubmedId":"","doi":"10.7554/eLife.86336"},{"reference":"Li Q, Stavropoulos N. 2016. Evaluation of Ligand-Inducible Expression Systems for Conditional Neuronal Manipulations of Sleep in Drosophila. G3 Genes|Genomes|Genetics. 6: 3351.","pubmedId":"","doi":"10.1534/g3.116.034132"},{"reference":"Li Y, Chouhan NS, Zhang SL, Moore RS, Noya SB, Shon J, Yue Z, Sehgal A. 2024. Modulation of RNA processing genes during sleep-dependent memory. eLife. 12: RP89023.","pubmedId":"","doi":"10.7554/eLife.89023.4"},{"reference":"Li Y, Haynes P, Zhang SL, Yue Z, Sehgal A. 2023. Ecdysone acts through cortex glia to regulate sleep in Drosophila. eLife. 12: e81723.","pubmedId":"","doi":"10.7554/eLife.81723"},{"reference":"Li Y, Zhou Z, Zhang X, Tong H, Li P, Zhang ZC, et al., Han J. 2013. Drosophila Neuroligin 4 Regulates Sleep through Modulating GABA Transmission. The Journal of Neuroscience. 33: 15545.","pubmedId":"","doi":"10.1523/JNEUROSCI.0819-13.2013"},{"reference":"Ly S, Lee DA, Strus E, Prober DA, Naidoo N. 2020. Evolutionarily Conserved Regulation of Sleep by the Protein Translational Regulator PERK. Current Biology. 30: 1639.","pubmedId":"","doi":"10.1016/j.cub.2020.02.030"},{"reference":"Osterwalder T, Yoon KS, White BH, Keshishian H. 2001. A conditional tissue-specific transgene expression system using inducible GAL4. Proceedings of the National Academy of Sciences. 98: 12596.","pubmedId":"","doi":"10.1073/pnas.221303298"},{"reference":"Persons JL, Abhilash L, Lopatkin AJ, Roelofs A, Bell EV, Fernandez MP, Shafer OT. 2022. PHASE: An Open-Source Program for the Analysis of Drosophila Ph ase, A ctivity, and S leep Under E ntrainment. Journal of Biological Rhythms. 37: 455.","pubmedId":"","doi":"10.1177/07487304221093114"},{"reference":"Pyfrom ES, Beveridge C, Haynes PR, Kanigicherla VA, Randolph CE, Carvalho Costa P, et al., Sehgal A. 2025. Neutral lipid processing in glia is sexually dimorphic and promotes sleep through diacylglycerol catabolism.","pubmedId":"","doi":"10.1101/2025.09.10.674993"},{"reference":"Ryder E, Blows F, Ashburner M, Bautista Llacer R, Coulson D, Drummond J, et al., Russell S. 2004. The DrosDel Collection. Genetics. 167: 797.","pubmedId":"","doi":"10.1534/genetics.104.026658"},{"reference":"Szabo, Vincze V, Chhatre AS, Jipa A, Bognar S, Varga KE, et al., Juhasz G. 2023. LC3-associated phagocytosis promotes glial degradation of axon debris after injury in Drosophila models. Nature Communications. 14: 3077.","pubmedId":"","doi":"10.1038/s41467-023-38755-4"},{"reference":"Tabuchi M, Lone SR, Liu S, Liu Q, Zhang J, Spira AP, Wu MN. 2015. Sleep Interacts with Aβ to Modulate Intrinsic Neuronal Excitability. Current Biology. 25: 702.","pubmedId":"","doi":"10.1016/j.cub.2015.01.016"},{"reference":"Vincze V, Eskudt Z, Feher Juhasz E, Chhatre AS, Jipa A, Galambos AR, et al., Szabo. 2026. Selective autophagy fine-tunes Stat92E activity by degrading Su(var)2-10/PIAS in Drosophila glia. Life Science Alliance. 9: e202503375.","pubmedId":"","doi":"10.26508/lsa.202503375"},{"reference":"Wu MN, Ho K, Crocker A, Yue Z, Koh K, Sehgal A. 2009. The Effects of Caffeine on Sleep in Drosophila Require PKA Activity, But Not the Adenosine Receptor. The Journal of Neuroscience. 29: 11029.","pubmedId":"","doi":"10.1523/JNEUROSCI.1653-09.2009"}],"title":"Drosophila pan-glial inducible Gal4 line alters baseline sleep
","reviews":[],"curatorReviews":[{"curator":{"displayName":"FlyBase Curators"},"openAcknowledgement":false,"submitted":null}]}]}},"species":{"species":[{"value":"acer saccharum","label":"Acer saccharum","imageSrc":"","imageAlt":"","mod":"TreeGenes","modLink":"https://treegenesdb.org","linkVariable":""},{"value":"achillea millefolium","label":"Achillea millefolium","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"acinetobacter baylyi","label":"Acinetobacter baylyi","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"actinobacteria bacterium","label":"Actinobacteria bacterium","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"adelges tsugae","label":"Adelges tsugae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"adenocaulon chilense","label":"Adenocaulon chilense","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"aedes japonicus","label":"Aedes japonicus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"aegorhinus vitulus","label":"Aegorhinus vitulus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"alaimidae","label":"Alaimidae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"allobates femoralis","label":"Allobates femoralis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"alnus glutinosa","label":"Alnus glutinosa","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"alosa aestivalis","label":"Alosa aestivalis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"alosa pseudoharengus","label":"Alosa pseudoharengus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"alternaria alternata","label":"Alternaria alternata","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"amynthas agrestis","label":"Amynthas Agrestis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ancylostoma caninum","label":"Ancylostoma caninum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ancylostoma ceylanicum","label":"Ancylostoma ceylanicum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"anemone multifida","label":"Anemone multifida","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"anguilla rostrata","label":"Anguilla rostrata","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"anisakis simplex","label":"Anisakis simplex","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"anomala albopilosa","label":"Anomala albopilosa","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"anthomyiidae sp","label":"Anthomyiidae sp","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"anthomyiidae sp","label":"Anthomyiidae sp","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"arabidopsis","label":"Arabidopsis","imageSrc":"arabidopsis.png","imageAlt":"Arabidopsis graphic by Zoe Zorn CC BY 4.0","mod":"TAIR","modLink":"https://arabidopsis.org","linkVariable":""},{"value":"architeuthis dux","label":"Architeuthis dux","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"arion vulgaris","label":"Arion vulgaris","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"armeria","label":"Armeria","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"artemia","label":"Artemia","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"arthrobacter sp.","label":"Arthrobacter sp.","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ascaridia","label":"Ascaridia","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ascaridia galli","label":"Ascaridia galli","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"asparagopsis taxiformis","label":"Asparagopsis taxiformis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"astatotilapia burtoni","label":"Astatotilapia burtoni","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"avena sativa","label":"Avena sativa","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"aves","label":"Aves","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bacillus","label":"Bacillus (firmicutes)","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bacillus cereus","label":"Bacillus cereus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bacillus mycoides","label":"Bacillus mycoides","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bacillus subtilis","label":"Bacillus subtilis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bacillus thuringiensis","label":"Bacillus thuringiensis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bacillus toyonensis","label":"Bacillus toyonensis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bacillus wiedmannii","label":"Bacillus wiedmannii","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bacteria","label":"Bacteria","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bacteriophage","label":"Bacteriophage","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bactrocera","label":"Bactrocera sp.","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"batrachospermum gelatinosum","label":"Batrachospermum gelatinosum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"betula lenta","label":"Betula lenta","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"betula nigra","label":"Betula nigra","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bombus dahlbohmii","label":"Bombus dahlbohmii","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bombus terrestris","label":"Bombus terrestris","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bombyx mori","label":"Bombyx mori","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bos taurus","label":"Bos Taurus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"brachygobius doriae","label":"Brachygobius doriae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"brassica oleracea","label":"Brassica oleracea","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"brassica rapa","label":"Brassica rapa","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"brugia malayi","label":"Brugia malayi","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"burkholderia thailandensis","label":"Burkholderia thailandensis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"buttiauxella","label":"Buttiauxella","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"caenorhabditis brenneri","label":"Caenorhabditis brenneri","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"caenorhabditis briggsae","label":"Caenorhabditis briggsae","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"c. elegans","label":"Caenorhabditis elegans","imageSrc":"c-elegans.jpg","imageAlt":"C. elegans graphic by Zoe Zorn CC BY 4.0","mod":"WormBase","modLink":"https://wormbase.org","linkVariable":""},{"value":"caenorhabditis inopinata","label":"Caenorhabditis inopinata","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"caenorhabditis japonica","label":"Caenorhabditis japonica","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"caenorhabditis nigoni","label":"Caenorhabditis nigoni","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"caenorhabditis remanei","label":"Caenorhabditis remanei","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"caenorhabditis tropicalis","label":"Caenorhabditis tropicalis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"calidifontibacillus","label":"Calidifontibacillus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"calidifontibacillus erzuremensis","label":"Calidifontibacillus erzuremensis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"calliphora sp","label":"Calliphora sp","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"caltha sagittata","label":"Caltha sagittata","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"cambarus latimanus","label":"Cambarus latimanus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"candida albicans","label":"Candida albicans","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"canis familiaris","label":"Canis familiaris","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"cannabis sativa","label":"Cannabis sativa","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"caretta caretta","label":"Caretta caretta","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"cassiopea xamachana","label":"Cassiopea xamachana","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"caulobacter vibrioides","label":"Caulobacter vibrioides","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"cephalopods","label":"Cephalopoda","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"cerastium arvense","label":"Cerastium arvense","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ceriodaphnia","label":"Ceriodaphnia","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ceroglossus suturalis","label":"Ceroglossus suturalis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"chaetoceros","label":"Chaetoceros","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"chamaecrista fasciculata","label":"Chamaecrista fasciculata","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"chilicola chalcidiformis","label":"Chilicola chalcidiformis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"chitinimonas","label":"Chitinimonas","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"chlamydomonas reinhardtii","label":"Chlamydomonas reinhardtii","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"chromobacterium","label":"Chromobacterium","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"chrysemys picta","label":"Chrysemys picta","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"chrysoperla rufilabris","label":"Chrysoperla rufilabris","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"citrus","label":"Citrus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"clavibacter sp.","label":"Clavibacter sp.","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"colinus virginianus","label":"Colinus virginianus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"crassostrea virginica","label":"Crassostrea virginica","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"crithidia fasciculata","label":"Crithidia fasciculata","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"cutibacterium acnes","label":"Cutibacterium acnes","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"cyanobacteria","label":"Cyanobacteria","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"daphnia","label":"Daphnia","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"daphnia pulex","label":"Daphnia pulex","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"diabrotica virgifera","label":"Diabrotica virgifera","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"diabrotica virgifera virgifera virus 1","label":"Diabrotica virgifera virgifera virus 1","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"d. discoideum","label":"Dictyostelium discoideum","imageSrc":"dicty.png","imageAlt":"D. discoideum","mod":"dictyBase","modLink":"http://dictybase.org","linkVariable":""},{"value":"diptera","label":"Diptera","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"dotocryptus bellicosus","label":"Dotocryptus bellicosus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"drechmeria coniospora","label":"Drechmeria coniospora","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"drosophila","label":"Drosophila","imageSrc":"drosophila.png","imageAlt":"Drosophila graphic by Zoe Zorn CC BY 4.0","mod":"FlyBase","modLink":"https://flybase.org/doi/","linkVariable":"doi"},{"value":"dryopteris campyloptera","label":"Dryopteris campyloptera","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"dryopteris expansa","label":"Dryopteris expansa","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"dryopteris intermedia","label":"Dryopteris intermedia","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"dugesia dorotocephala","label":"Dugesia dorotocephala","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"elasmobranchii","label":"Elasmobranchii","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"embryophyta","label":"Embryophyta","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"enoploteuthis chunii","label":"Enoploteuthis chunii","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"enterobacter aerogenes","label":"Enterobacter aerogenes","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"enterococcus raffinosus","label":"Enterococcus raffinosus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"epichloë coenophiala","label":"Epichloë coenophiala","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"equus caballus","label":"Equus caballus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"erigeron sp","label":"Erigeron sp","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"eristalis","label":"Eristalis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"eruca vesicaria","label":"Eruca vesicaria","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"erwinia carotovora","label":"Erwinia carotovora","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"erythronium americanum","label":"Erythronium americanum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"escherichia coli","label":"Escherichia coli","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"eukaryota","label":"Eukaryotes","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"felis catus","label":"Felis catus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"francisella novicida","label":"Francisella novicida","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"francisella tularensis","label":"Francisella tularensis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"fraxinus americana","label":"Fraxinus americana","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"fucus distichus","label":"Fucus distichus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"fungi","label":"Fungi","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"gasteropelecus sp.","label":"Gasteropelecus sp.","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"geranium sp","label":"Geranium sp","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"girardia","label":"Girardia","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"glaucomys volans","label":"Glaucomys volans","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"glycine max","label":"Glycine max","imageSrc":"","imageAlt":"","mod":"Soybase","modLink":"https://soybase.org","linkVariable":""},{"value":"glyptemys insculpta","label":"Glyptemys insculpta","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"gossypium hirsutum","label":"Gossypium hirsutum","imageSrc":"","imageAlt":"","mod":"CottonGen","modLink":"https://www.cottongen.org/","linkVariable":""},{"value":"gromphadorhina portentosa","label":"Gromphadorhina portentosa","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"gryllodes sigillatus","label":"Gryllodes sigillatus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"haliotis rufescens","label":"Haliotis rufescens","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"hepacivirus hominis","label":"Hepatitis C Virus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"herpes simplex virus type 1","label":"Herpes simplex virus type 1","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"human","label":"Human","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"human coronavirus oc43","label":"Human coronavirus OC43","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"hydra vulgaris","label":"Hydra vulgaris","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"hydropsyche sp","label":"Hydropsyche sp","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"hymenoptera","label":"Hymenoptera","imageSrc":"","imageAlt":"","mod":"Hymenoptera Genome Database","modLink":"https://hymenoptera.elsiklab.missouri.edu/","linkVariable":""},{"value":"hypochaeris radicata","label":"Hypochaeris radicata","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"hypodynerus vespiformis","label":"Hypodynerus vespiformis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"iflaviridae","label":"Iflaviridae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"iflavuris","label":"Iflavirus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ipomoea hederacea","label":"Ipomoea hederacea","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ischnomera","label":"Ischnomera","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ischnomera ruficollis","label":"Ischnomera ruficollis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"julidochromis marlieri","label":"Julidochromis marlieri","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"juniperus virginiana","label":"Juniperus virginiana","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"kluyveromyces marxianus","label":"Kluyveromyces marxianus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"l. casei","label":"L. casei","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"lacticaseibacillus casei","label":"Lacticaseibacillus casei","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"larentiinae sp","label":"Larentiinae sp","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"laurus nobilis","label":"Laurus nobilis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"lepidoptera","label":"Lepidoptera","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"leucanthemum vulgare","label":"Leucanthemum vulgare","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"linepithema humile","label":"Linepithema humile","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"liometopum occidentale","label":"Liometopum occidentale","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"lolium arundinaceum","label":"Lolium arundinaceum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"lumbriculus variegatus","label":"Lumbriculus variegatus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"lumbricus terrestris","label":"Lumbricus terrestris","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"lupinus polyphyllus","label":"Lupinus polyphyllus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"lycorma delicatula","label":"Lycorma delicatula","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"lynx rufus","label":"Lynx rufus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"magnaporthe oryzae","label":"Magnaporthe oryzae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"mammalia","label":"Mammalia","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"manihot esculenta","label":"Manihot esculenta","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"medicago lupulina","label":"Medicago lupulina","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"meloidogyne","label":"Meloidogyne","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"mimus polyglottos","label":"Mimus polyglottos","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"bryophyta","label":"Mosses","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"mouse","label":"Mouse","imageSrc":"","imageAlt":"","mod":"MGI","modLink":"https://informatics.jax.org","linkVariable":""},{"value":"m. minutoides","label":"Mus minutoides","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"mycobacterium smegmatis","label":"Mycobacterium smegmatis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"nakaseomyces glabratus","label":"Nakaseomyces glabratus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"nauphoeta cinerea","label":"Nauphoeta cinerea","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"neurospora","label":"Neurospora","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"n. benthamiana","label":"Nicotiana benthamiana","imageSrc":"","imageAlt":"","mod":"Solgenomics Network","modLink":"https://solgenomics.net/organism/Nicotiana_benthamiana/genome","linkVariable":""},{"value":"nicotiana tabacum","label":"Nicotiana tabacum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"noctuidae","label":"Noctuidae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"noctuidae sp","label":"Noctuidae sp","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"nothobranchius furzeri","label":"Nothobranchius furzeri","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"onchocerca volvulus","label":"Onchocerca volvulus","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"orconectes virilis","label":"Orconectes virilis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ormia ochracea","label":"Ormia ochracea","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"o. sativa","label":"Oryza sativa","imageSrc":"","imageAlt":"","mod":"Gramene","modLink":"https://www.gramene.org/","linkVariable":""},{"value":"other","label":"Other","imageSrc":"","imageAlt":"","mod":null,"modLink":null,"linkVariable":null},{"value":"oxalis enneaphylla","label":"Oxalis enneaphylla","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"paenarthrobacter nicotinovorans","label":"Paenarthrobacter nicotinovorans","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"paenarthrobacter nicotinovorans","label":"Paenarthrobacter nicotinovorans","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pantoea","label":"Pantoea","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pantoea agglomerans","label":"Pantoea agglomerans","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"papaver sp","label":"Papaver sp","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"paramecium bursaria","label":"Paramecium bursaria","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"partitiviridae","label":"Partitiviridae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pelodiscus sinensis","label":"Pelodiscus sinensis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"perezia recurvata","label":"Perezia recurvata","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"petromyzon marinus","label":"Petromyzon marinus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"photinus pyralis","label":"Photinus pyralis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"photinus pyralis associated partiti-like virus","label":"Photinus pyralis associated partiti-like virus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"photinus pyralis iflavirus 1","label":"Photinus pyralis iflavirus 1","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"physcomitrium patens","label":"Physcomitrium patens","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pinus strobus","label":"Pinus strobus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pinus taeda","label":"Pinus taeda","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"platycheirus","label":"Platycheirus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"plectus sambesii","label":"Plectus sambesii","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pogonomyrmex occidentalis","label":"Pogonomyrmex occidentalis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"poncirus trifoliata","label":"Poncirus trifoliata","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"populus deltoides","label":"Populus deltoides","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"potato virus y","label":"Potato virus Y","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"primula magellanica","label":"Primula magellanica","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pristionchus pacificus","label":"Pristionchus pacificus","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"prunus persica","label":"Prunus persica","imageSrc":"","imageAlt":"","mod":"Genome Database for Rosaceae","modLink":"https://www.rosaceae.org/","linkVariable":""},{"value":"psalmopoeus iriminia","label":"Psalmopoeus iriminia","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pseudanabaena sp.","label":"Pseudanabaena sp.","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pseudomonas","label":"Pseudomonas","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pseudomonas aeruginosa","label":"Pseudomonas aeruginosa","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pseudomonas glycinae","label":"Pseudomonas glycinae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pseudomonas putida","label":"Pseudomonas putida","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pseudomonas syringae","label":"Pseudomonas syringae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"pterophyllum scalare","label":"Pterophyllum scalare","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"python regius","label":"Python regius","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"quercus macrocarpa","label":"Quercus macrocarpa","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ralstonia solanacearum","label":"Ralstonia solanacearum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ranitomeya imitator","label":"Ranitomeya imitator","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ranunculus peduncularis","label":"Ranunculus peduncularis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"rat","label":"Rat","imageSrc":"","imageAlt":"","mod":"RGD","modLink":"https://rgd.mcw.edu","linkVariable":""},{"value":"rheinheimera","label":"Rheinheimera","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ribes rubrum","label":"Ribes rubrum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"sars-cov-2","label":"SARS-CoV-2","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"s. cerevisiae","label":"Saccharomyces cerevisiae","imageSrc":"yeast.png","imageAlt":"Yeast graphic by Zoe Zorn CC BY 4.0","mod":"SGD","modLink":"https://yeastgenome.org","linkVariable":""},{"value":"saccharomyces paradoxus","label":"Saccharomyces paradoxus ","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"s. uvarum","label":"Saccharomyces uvarum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"schistosoma","label":"Schistosoma","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"schizosaccharomyces japonicus","label":"Schizosaccharomyces japonicus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"s. pombe","label":"Schizosaccharomyces pombe","imageSrc":"pombe.png","imageAlt":"Pombe graphic by Zoe Zorn © Caltech","mod":"PomBase","modLink":"https://www.pombase.org/reference/PMID:","linkVariable":"pmId"},{"value":"schmidtea mediterranea","label":"Schmidtea mediterranea","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"senecio sp","label":"Senecio sp","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"simocephalus","label":"Simocephalus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"siraitia grosvenorii","label":"Siraitia grosvenorii","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"solanum lycopersicum","label":"Solanum lycopersicum","imageSrc":"","imageAlt":"","mod":"Solgenomics Network","modLink":"https://solgenomics.net/organism/1/view/","linkVariable":""},{"value":"sorghum","label":"Sorghum","imageSrc":"","imageAlt":"","mod":"SorghumBase","modLink":"https://www.sorghumbase.org","linkVariable":""},{"value":"spiroplasma eriocheiris","label":"Spiroplasma eriocheiris","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"staphylococcus aureus","label":"Staphylococcus aureus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"staphylococcus epidermidis","label":"Staphylococcus epidermidis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"steinernema carpocapsae","label":"Steinernema carpocapsae","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"https://wormbase.org","linkVariable":""},{"value":"steinernema hermaphroditum","label":"Steinernema hermaphroditum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"stenotrophomonas geniculata","label":"Stenotrophomonas geniculata","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"streptococcus gordonii ","label":"Streptococcus gordonii ","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"streptococcus mutans","label":"Streptococcus mutans","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":" streptococcus pneumoniae","label":"Streptococcus pneumoniae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"s. purpuratus","label":"Strongylocentrotus purpuratus","imageSrc":"","imageAlt":"","mod":"Echinobase","modLink":"https://www.echinobase.org","linkVariable":""},{"value":"strongyloides ratti","label":"Strongyloides ratti","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"sulfolobus","label":"Sulfolobus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"symphoricarpos albus","label":"Symphoricarpos albus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"syncirsodes","label":"Syncirsodes","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"synechococcus elongatus","label":"Synechococcus elongatus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"syrphidae","label":"Syrphidae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"tarantobelus jeffdanielsi","label":"Tarantobelus jeffdanielsi","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"taraxacum officinale","label":"Taraxacum officinale","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"tatochila theodice","label":"Tatochila theodice","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"tetrahymena","label":"Tetrahymena","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"tetramorium immigrans","label":"Tetramorium immigrans","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"tomato brown rugose fruit virus","label":"ToBRFV","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"trachemys scripta","label":"Trachemys scripta","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"tribolium castaneum","label":"Tribolium castaneum","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"trichoptera","label":"Trichoptera","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"trichuris muris","label":"Trichuris muris","imageSrc":"","imageAlt":"","mod":"WormBase","modLink":"www.wormbase.org","linkVariable":""},{"value":"trifolium repens","label":"Trifolium repens","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"trypoxylus dichotomus","label":"Trypoxylus dichotomus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"tsuga canadensis","label":"Tsuga canadensis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"ulva expansa","label":"Ulva expansa","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"universal","label":"Universal","imageSrc":"","imageAlt":"","mod":null,"modLink":null,"linkVariable":null},{"value":"vargula hilgendorfii","label":"Vargula hilgendorfii","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"vespula vulgaris","label":"Vespula vulgaris","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"virus","label":"Virus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"watasenia scintillans","label":"Watasenia scintillans","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"wolbachia pipientis","label":"Wolbachia pipientis","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"xenopus","label":"Xenopus","imageSrc":"xenopus.png","imageAlt":"Xenopus graphic by Zoe Zorn CC BY 4.0","mod":"XenBase","modLink":"https://xenbase.org","linkVariable":""},{"value":"xenorhabdus griffiniae","label":"Xenorhabdus griffiniae","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"yramea cytheris","label":"Yramea cytheris","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"zaprionus indianus","label":"Zaprionus indianus","imageSrc":"","imageAlt":"","mod":"","modLink":"","linkVariable":""},{"value":"zea mays","label":"Zea mays","imageSrc":"","imageAlt":"","mod":"MaizeGDB","modLink":"https://www.maizegdb.org","linkVariable":""},{"value":"zebrafish","label":"Zebrafish","imageSrc":"zebrafish.png","imageAlt":"Zebrafish graphic by Zoe Zorn CC BY 4.0","mod":"ZFIN","modLink":"https://zfin.org","linkVariable":""}]}},"pageContext":{"id":"d6ddc7f7-4047-4da3-bae8-976917793a2a","citedBy":[],"parsedCsv":{"csvHeader":[],"csvData":[]}}}, "staticQueryHashes": ["2114697108"]}