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.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $18.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
NCBI Reference Sequence
We are grateful to A. Fire (Stanford University) for sharing unpublished results and to C.T. Hittinger (University of Wisconsin-Madison), D. Sieburth (University of Southern California), E.M. Jorgensen (University of Utah), C.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.
Integrated supplementary information
Blue light induces paralysis in transgenic animals carrying a GABAergic driver and a channelrhodopsin (ChR2) effector
No response to blue light in transgenic animals only carrying a channelrhodopsin (ChR2) effector
Calcium imaging in body wall muscles of animals carrying a body wall muscle driver and a GCaMP6s::SL2::mKate2 effector
Expressing a histamine-gated chloride channel HisCl1 in body wall muscle induces flaccid paralysis on histamine plates. Worms with either the driver or the effector alone fail to respond to histamine
Expressing a tetanus toxin light chain (TeTx) in GABAergic neurons blocks neurotransmission and leads to the characteristic “shrink” phenotype. Transgenic worms with either the driver or the effector don't display the “shrink” phenotype