A toolkit for GFP-mediated tissue-specific protein degradation in C. elegans

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Development. 2017 Jul;144:2694-2701. doi: 10.1242/dev.150094

A toolkit for GFP-mediated tissue-specific protein degradation in C. elegans Wang (Shaohe) Lab

Wang S, Tang NH, Lara-Gonzalez P, Zhao Z, Cheerambathur DK, Prevo B, Chisholm AD, Desai A, Oegema K

Note: Research in this publication was not performed at Janelia.

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Abstract

Proteins that are essential for embryo production, cell division and early embryonic events are frequently reused 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 uses 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 with 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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A toolkit for GFP-mediated tissue-specific protein degradation in C. elegans

Note: Research in this publication was not performed at Janelia.

Abstract

Janelia Authors