Repurposing an endogenous degradation system for rapid and targeted depletion of C. elegans proteins
Open accessAbstract
The capability to conditionally inactivate gene function is essential for understanding the molecular basis of development. In gene and mRNA targeting approaches, protein products can perdure, complicating genetic analysis. Current methods for selective protein degradation require drug treatment or take hours for protein removal, limiting their utility in studying rapid developmental processes in vivo. Here, we repurpose an endogenous protein degradation system to rapidly remove targeted C. elegans proteins. We show that upon expression of the E3 ubiquitin ligase substrate-recognition subunit ZIF-1, proteins tagged with the ZF1 zinc-finger domain can be quickly degraded in all somatic cell types examined with temporal and spatial control. We demonstrate that genes can be engineered to become conditional loss-of-function alleles by introducing sequences encoding the ZF1 tag into endogenous loci. Finally, we use ZF1 tagging to establish the site of cdc-42 gene function during a cell invasion event. ZF1 tagging provides a powerful new tool for the analysis of dynamic developmental events.
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Tagging POIs with a ZF1 motif enables their recruitment and degradation via endogenous CRL machinery.
“By tagging POIs with such a ZF1 motif, it is possible to recruit and degrade them via endogenous CRL machinery [52].”
From Advances in targeted degradation of endogenous proteins (2019) · cites this paper as [52]
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Differential adhesion regulates neurite placement via a retrograde zippering mechanismPreprint
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- Authors
- Armenti, S. T., Lohmer, L. L., Sherwood, D. R., & Nance, J.
- Affiliations
- NYU Langone Health (US)
- Duke University (US)
- Corresponding author
- Nance, Jeremy
- Venue
- Development
- Publisher
- The Company of Biologists
- Published
- 2014-12-01
- Volume
- 141
- Issue
- 23
- Pages
- 4640-4647
- ISSN
- 0950-1991 (ISSN-L)
- 1477-9129
- PMC
- PMC4302935
- OpenAlex
- W2137117135
- Cited by
- 124
- References
- 65
- Citations by year
- 2026: 10 · 2025: 4 · 2024: 11 · 2023: 19 · 2022: 13 · 2021: 17 · 2020: 9 · 2019: 12 · 2018: 9 · 2017: 6 · 2016: 10 · 2015: 4
- FWCI
- 2.74
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Development (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- HighWire Press Open Archive (submittedVersion)
- Funders
- National Institute of General Medical Sciences (R01GM079320, R01GM078341, T32GM007308, R01GM098492, R01GM100083)
- National Institute of Diabetes and Digestive and Kidney Diseases (F30DK093197)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
- Keywords
- protein degradation, E3 ubiquitin ligase, conditional alleles, Cdc42, cell invasion, Caenorhabditis elegans, developmental dynamics
- MeSH terms
- Animals, Carrier Proteins, Fluorescence, Gene Expression Regulation, Genetic Engineering, Image Processing, Computer-Assisted, Kinetics, Plasmids, Blotting, Western, Zinc Fingers, Caenorhabditis elegans, DNA Primers, Transgenes, Caenorhabditis elegans Proteins, Ubiquitin-Protein Ligases, Proteolysis
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