RRC ID 51413
Author Wang S, Tang NH, Lara-Gonzalez P, Zhao Z, Cheerambathur DK, Prevo B, Chisholm AD, Desai A, Oegema K.
Title A toolkit for GFP-mediated tissue-specific protein degradation in C. elegans.
Journal Development
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.
Volume 144(14)
Pages 2694-2701
Published 2017-7-15
DOI 10.1242/dev.150094
PII dev.150094
PMID 28619826
PMC PMC5536931
MeSH Animals Animals, Genetically Modified Axons / metabolism Caenorhabditis elegans / genetics Caenorhabditis elegans / growth & development Caenorhabditis elegans / metabolism* Caenorhabditis elegans Proteins / genetics Caenorhabditis elegans Proteins / metabolism Carrier Proteins / genetics Carrier Proteins / metabolism Genes, Helminth Genetic Techniques Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism* MAP Kinase Kinase Kinases / genetics MAP Kinase Kinase Kinases / metabolism Mutation Proteolysis Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Single-Domain Antibodies / genetics Single-Domain Antibodies / metabolism Tissue Distribution
IF 5.611
Times Cited 24
C.elegans tm4024