Xiao, Yutong, Yee, Callista, Martinez, Michael, Zhao, Chris, Zhang, Wan, Shen, Kang, Matus, David, Hammell, Christopher (2022) An expandable FLP-ON::TIR1 system for precise spatiotemporal protein degradation in C. elegans. bioRxiv. (Submitted)
Abstract
The auxin-inducible degradation system has been widely adopted in the C. elegans research community for its ability to empirically control the spatiotemporal expression of target proteins. This system can efficiently degrade auxin-inducible degron (AID)-tagged proteins via the expression of a ligand-activatable AtTIR1 protein derived from A. thaliana that adapts target proteins to the endogenous C. elegans proteosome. While broad expression of AtTIR1 using strong, ubiquitous promoters can lead to rapid degradation of AID-tagged proteins, cell type-specific expression of AtTIR1 using spatially restricted promoters often results in less efficient target protein degradation. To circumvent this limitation, we have developed a FLP/FRT3-based system that functions to reanimate a dormant, high-powered promoter that can drive sufficient AtTIR1expression in a cell type-specific manner. We benchmark the utility of this system by generating a number of tissue specific FLP-ON::TIR1 drivers to reveal genetically separable cell type-specific phenotypes for several target proteins. We also demonstrate that the FLP-ON::TIR1 system is compatible with enhanced degron epitopes. Finally, we provide an expandable toolkit utilizing the basic FLP-ON::TIR1 system that can be adapted to drive optimized AtTIR1expression in any tissue or cell type of interest.
| Item Type: | Paper |
|---|---|
| Subjects: | organism description > animal > C elegans |
| CSHL Authors: | |
| Communities: | CSHL labs > Hammell C. lab |
| SWORD Depositor: | CSHL Elements |
| Depositing User: | CSHL Elements |
| Date: | 2022 |
| Date Deposited: | 29 Sep 2023 15:59 |
| Last Modified: | 27 Dec 2023 19:32 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/41064 |
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