A toolkit for GFP-mediated tissue-specific protein degradation in C. elegans [TECHNIQUES AND RESOURCES REPORT]

Shaohe Wang, Ngang Heok Tang, Pablo Lara-Gonzalez, Zhiling Zhao, Dhanya K. Cheerambathur, Bram Prevo, Andrew D. Chisholm, Arshad Desai, and Karen Oegema

Proteins essential for embryo production, cell division, and early embryonic events are frequently re-utilized later in embryogenesis, during organismal development, or in the adult. Examining protein function across these different biological contexts requires tissue-specific perturbation. Here, we describe a method that utilizes expression of a fusion between a GFP-targeting nanobody and a SOCS-box containing ubiquitin ligase adaptor to target GFP tagged proteins for degradation. When combined with endogenous locus GFP tagging by CRISPR-Cas9 or rescue of a null mutant with a GFP fusion, this approach enables routine and efficient tissue-specific protein ablation. We show that this approach works in multiple tissues—the epidermis, intestine, body wall muscle, ciliated sensory neurons, and touch receptor neurons—where it recapitulates expected loss-of-function mutant phenotypes. The transgene toolkit and the strain set described here will complement existing approaches to enable routine analysis of the tissue-specific roles of C. elegans proteins.

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