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cGAL, a temperature-robust GAL4–UAS system for Caenorhabditis elegans

Wang, Han and Liu, Jonathan and Gharib, Shahla and Chai, Cynthia M. and Schwarz, Erich M. and Pokala, Navin and Sternberg, Paul W. (2017) cGAL, a temperature-robust GAL4–UAS system for Caenorhabditis elegans. Nature Methods, 14 (2). pp. 145-148. ISSN 1548-7091. PMCID PMC5693259. https://resolver.caltech.edu/CaltechAUTHORS:20161117-125604575

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[img] Image (JPEG) (Supplementary Figure 2: Performance of different DBDs from Gal4 proteins at room temperature) - Supplemental Material
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[img] Image (JPEG) (Supplementary Figure 3: Functional verification of integrated effectors) - Supplemental Material
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[img] Video (MPEG) (Video 1: Functional verification of a ChR2 effector with a GABAergic driver) - Supplemental Material
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[img] Video (MPEG) (Video 2: Negative control of a ChR2 effector without driver) - Supplemental Material
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[img] Video (MPEG) (Video 3: Functional verification of a GCaMP6s effector with a body wall muscle driver) - Supplemental Material
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[img] Video (MPEG) (Video 4: Functional verification of a HisCl1 effector with a body wall muscle driver) - Supplemental Material
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[img] Video (MPEG) (Video 5: Functional verification of a TeTx effector with a GABAergic driver) - Supplemental Material
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Abstract

The GAL4–UAS system is a powerful tool for manipulating gene expression, but its application in Caenorhabditis elegans has not been described. Here we systematically optimize the system's three main components to develop a temperature-optimized GAL4–UAS system (cGAL) that robustly controls gene expression in C. elegans from 15 to 25 °C. We demonstrate this system's utility in transcriptional reporter analysis, site-of-action experiments and exogenous transgene expression; and we provide a basic driver and effector toolkit.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1038/nmeth.4109DOIArticle
http://rdcu.be/nYvdPublisherFree ReadCube access
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5693259PubMed CentralArticle
ORCID:
AuthorORCID
Wang, Han0000-0002-1933-5762
Schwarz, Erich M.0000-0003-3151-4381
Sternberg, Paul W.0000-0002-7699-0173
Additional Information:© 2016 Macmillan Publishers Limited. Received 22 May; Accepted 18 November; Published online 19 December 2016. We are grateful to A. Fire (Stanford University) for sharing unpublished results and to C.T. Hittinger (University of Wisconsin-Madison), D. Sieburth (University of Southern California), E.M. Jorgensen (University of Utah), C. Bargmann (Rockefeller University) and A. Fire (Stanford University) for reagents. We thank H. Korswagen (Hubrecht Institute) for thoughtful discussion. N.P. thanks C. Bargmann for her support. We thank M. Bao, Y.M. Kim, D. Leighton, J. DeModena and G. Medina for technical assistance; and WormBase for technical support. We also thank M. Kato, H. Schwartz, D. Angeles-Albores and other members of the Sternberg lab for editorial comments on the manuscript. Some strains were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (grant P40 OD010440). Some imaging was performed at the Caltech Biological Imaging Facility with the support of the Caltech Beckman Institute and the Arnold and Mabel Beckman Foundation. H.W. is supported by the Della Martin Fellowship. J.L. was supported by NIH grant T32GM007616. This work is supported by the Howard Hughes Medical Institute, with which P.W.S. is an investigator. Data availability: The source data used for the plots in the main and supplementary figures are included in the supplementary materials. Other data are also available from the corresponding author upon request. The coding sequence of Gal4p from Saccharomyces kudriavzevii is in GenBank (GU299177.1). The plasmids for cGAL driver (pHW393, plasmid no. 85583) and effector (pHW394, plasmid no. 85584) are available from Addgene. Source data for Figures 1, 2, 3 and Supplementary Figures 1 and 2 are available online. Author Contributions: H.W., J.L. and P.W.S. conceived the project. H.W. and J.L. performed the experiments, analyzed the data and wrote the paper. S.G. helped with molecular cloning and strain handling. E.M.S. devised the idea of trying Gal4p from yeast species with lower growth temperatures. C.M.C. and N.P. contributed reagents. The authors declare no competing financial interests.
Funders:
Funding AgencyGrant Number
NIHP40 OD010440
Caltech Beckman InstituteUNSPECIFIED
Arnold and Mabel Beckman FoundationUNSPECIFIED
Della Martin FoundationUNSPECIFIED
NIH Predoctoral FellowshipT32GM007616
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Issue or Number:2
PubMed Central ID:PMC5693259
Record Number:CaltechAUTHORS:20161117-125604575
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20161117-125604575
Official Citation:cGAL, a temperature-robust GAL4–UAS system for Caenorhabditis elegans Han Wang, Jonathan Liu, Shahla Gharib, Cynthia M Chai, Erich M Schwarz, Navin Pokala & Paul W Sternberg Nature Methods (2017) doi:10.1038/nmeth.4109
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:72124
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:20 Dec 2016 00:59
Last Modified:29 Oct 2019 21:37

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