Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans

The auxin-inducible degron (AID) provides reversible, spatiotemporal control for the knockdown of target proteins. Here, we present a protocol for AID-mediated protein knockdown in Caenorhabditis elegans. We describe steps for generating the knock-in mutants using two CRISPR-Cas9 genome editing tech...

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Detalles Bibliográficos
Autores principales: Kurashina, Mizuki, Mizumoto, Kota
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9860162/
https://www.ncbi.nlm.nih.gov/pubmed/36640369
http://dx.doi.org/10.1016/j.xpro.2022.102028
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author Kurashina, Mizuki
Mizumoto, Kota
author_facet Kurashina, Mizuki
Mizumoto, Kota
author_sort Kurashina, Mizuki
collection PubMed
description The auxin-inducible degron (AID) provides reversible, spatiotemporal control for the knockdown of target proteins. Here, we present a protocol for AID-mediated protein knockdown in Caenorhabditis elegans. We describe steps for generating the knock-in mutants using two CRISPR-Cas9 genome editing techniques and preparing the auxin-containing nematode growth media (NGM) plates. We also detail AID-mediated spatiotemporal protein knockdown. For complete details on the use and execution of this protocol, please refer to Kurashina et al. (2021).(1)
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spelling pubmed-98601622023-01-22 Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans Kurashina, Mizuki Mizumoto, Kota STAR Protoc Protocol The auxin-inducible degron (AID) provides reversible, spatiotemporal control for the knockdown of target proteins. Here, we present a protocol for AID-mediated protein knockdown in Caenorhabditis elegans. We describe steps for generating the knock-in mutants using two CRISPR-Cas9 genome editing techniques and preparing the auxin-containing nematode growth media (NGM) plates. We also detail AID-mediated spatiotemporal protein knockdown. For complete details on the use and execution of this protocol, please refer to Kurashina et al. (2021).(1) Elsevier 2023-01-13 /pmc/articles/PMC9860162/ /pubmed/36640369 http://dx.doi.org/10.1016/j.xpro.2022.102028 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Kurashina, Mizuki
Mizumoto, Kota
Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans
title Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans
title_full Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans
title_fullStr Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans
title_full_unstemmed Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans
title_short Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans
title_sort targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in caenorhabditis elegans
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9860162/
https://www.ncbi.nlm.nih.gov/pubmed/36640369
http://dx.doi.org/10.1016/j.xpro.2022.102028
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