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

Shaohe Wang, Ngang Heok Tang, Pablo Lara-Gonzalez, Bram Prevo, Dhanya K. Cheerambathur, Andrew D. Chisholm, Arshad Desai, Karen Oegema
doi: https://doi.org/10.1101/104398
Shaohe Wang
1Ludwig Institute for Cancer Research, Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, California 92093, USA
2Present address: Cell Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892, USA
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Ngang Heok Tang
3Section of Neurobiology, Division of Biological Sciences, University of California San Diego, La Jolla, California 92093, USA
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Pablo Lara-Gonzalez
1Ludwig Institute for Cancer Research, Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, California 92093, USA
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Bram Prevo
1Ludwig Institute for Cancer Research, Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, California 92093, USA
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Dhanya K. Cheerambathur
1Ludwig Institute for Cancer Research, Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, California 92093, USA
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Andrew D. Chisholm
3Section of Neurobiology, Division of Biological Sciences, University of California San Diego, La Jolla, California 92093, USA
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Arshad Desai
1Ludwig Institute for Cancer Research, Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, California 92093, USA
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Karen Oegema
1Ludwig Institute for Cancer Research, Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, California 92093, USA
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  • For correspondence: koegema@ucsd.edu
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Abstract

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 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, sensory neurons, and touch 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.

Footnotes

  • Summary statement: An easy-to-implement protein degradation method that targets GFP fusions eliminates proteins in specific C. elegans tissues.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted January 30, 2017.
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A toolkit for tissue-specific protein degradation in C. elegans
Shaohe Wang, Ngang Heok Tang, Pablo Lara-Gonzalez, Bram Prevo, Dhanya K. Cheerambathur, Andrew D. Chisholm, Arshad Desai, Karen Oegema
bioRxiv 104398; doi: https://doi.org/10.1101/104398
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A toolkit for tissue-specific protein degradation in C. elegans
Shaohe Wang, Ngang Heok Tang, Pablo Lara-Gonzalez, Bram Prevo, Dhanya K. Cheerambathur, Andrew D. Chisholm, Arshad Desai, Karen Oegema
bioRxiv 104398; doi: https://doi.org/10.1101/104398

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