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Application of the red-shifted channelrhodopsin Chrimson for the Caenorhabditis elegans cGAL bipartite system

Cao, Mengyi and Chai, Cynthia and Liu, Jonathan and Sternberg, Paul W. (2018) Application of the red-shifted channelrhodopsin Chrimson for the Caenorhabditis elegans cGAL bipartite system. microPublication Biology . 10.17912/2JGW-FJ52. ISSN 2578-9430. PMCID PMC7282527. doi:10.17912/2JGW-FJ52.

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Channelrhodopsins are light-gated ion channels that serve as photoreceptors in photosynthetic microbes and have been applied as crucial optogenetic tools in genetic model organisms. When expressed in animals, they enable light-inducible control of ionic membrane permeability, which directly manipulates the activity of neurons expressing the protein. The application of channelrhodopsin-based optogenetics is particularly powerful when used in conjugation with the cGAL (GAL4-UAS) bipartite system (Wang, 2017). The mating of neuron-specific GAL4 driver lines to new channelrhodopsin effector lines could expand the genetic toolkit to perturb and manipulate neural circuits in the organism. Blue light-gated channelrhodopsins have been widely used in C. elegans neurobiology but often have to be performed in lite-1(ce314) mutant backgrounds because short-wavelength blue light is an aversive cue in wild-type animals and directly affects C. elegans neuronal physiology. Previously, a red light-gated variant of channelrhodopsin, termed Chrimson, has been successfully applied in Drosophila and mice, and has recently been codon-optimized for use in C. elegans (Klapoetke, 2014; Schild, 2015). Here, we constructed a Chrimson (15xUAS::chrimson::gfp) cGAL effector line. We introduced the UAS::chrimson::gfp effector DNA construct as an extrachromosomal array into a previously published cGAL pan-neuronal driver line (PS6961 syIs334) and generated integrants on chromosome II (PS8023, syIs503) and chromosome V (PS8024, syIs504) (Table 1) via standard X-ray irradiation. We showed Chrimson-GFP expression in the C. elegans head and tail neurons (Fig. 1A-1D). We also showed that red light could induce a seizure-like motility phenotype in C. elegans expressing Chrimson-GFP in a pan-neuronal manner (videos), while the negative controls expressing only the effector, or without light induction showed regular motility as expected (Table 2). The body curvature maps from normal and seizure-like motilities showed distinct patterns (Fig. 1E and 1F). We report the effector construct of red-light-gated channelrhodopsin Chrimson as an addition to our cGAL toolkit, which could be widely used in future research to overcome the technical restrictions of blue light-gated channelrhodopsins in C. elegans.

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Sternberg, Paul W.0000-0002-7699-0173
Additional Information:© 2018 by the authors. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: August 7, 2018; Accepted: August 21, 2018; Published: August 22, 2018. We thank Dr. Han Wang for his comments, suggestions, and support. Funding: R21 MH115454/MH/NIMH NIH HHS/United States
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NIHR21 MH115454
National Institute of Mental Health (NIMH)UNSPECIFIED
PubMed Central ID:PMC7282527
Record Number:CaltechAUTHORS:20200630-074613816
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Official Citation:Cao, M; Chai, C; Liu, J; Sternberg, PW (2018). Application of the red-shifted channel rhodopsin Chrimson for the Caenorhabditis elegans cGAL bipartite system. microPublication Biology. 10.17912/2JGW-FJ52. Download: RIS BibTeX
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:104154
Deposited By: Tony Diaz
Deposited On:30 Jun 2020 17:25
Last Modified:16 Nov 2021 18:28

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