Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans
Summary: 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 edi...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672200908X |
| _version_ | 1797952786880528384 |
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| author | Mizuki Kurashina Kota Mizumoto |
| author_facet | Mizuki Kurashina Kota Mizumoto |
| author_sort | Mizuki Kurashina |
| collection | DOAJ |
| description | Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-10T22:51:36Z |
| format | Article |
| id | doaj.art-70ae19c480524bf5b59f613614852e80 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-10T22:51:36Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-70ae19c480524bf5b59f613614852e802023-01-15T04:22:39ZengElsevierSTAR Protocols2666-16672023-03-0141102028Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegansMizuki Kurashina0Kota Mizumoto1Department of Zoology, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada; Graduate Program in Cell & Developmental Biology, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada; The Life Sciences Institute, The University of British Columbia, Vancouver, BC V6T 1Z3, CanadaDepartment of Zoology, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada; Graduate Program in Cell & Developmental Biology, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada; The Life Sciences Institute, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada; Djavad Mowafaghian Centre for Brain Health, The University of British Columbia, Vancouver, BC V6T 1Z3, Canada; Corresponding authorSummary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S266616672200908XCRISPRDevelopmental biologyGeneticsModel OrganismsNeuroscience |
| spellingShingle | Mizuki Kurashina Kota Mizumoto Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans STAR Protocols CRISPR Developmental biology Genetics Model Organisms Neuroscience |
| 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 | CRISPR Developmental biology Genetics Model Organisms Neuroscience |
| url | http://www.sciencedirect.com/science/article/pii/S266616672200908X |
| work_keys_str_mv | AT mizukikurashina targetingendogenousproteinsforspatialandtemporalknockdownusingauxininducibledegronincaenorhabditiselegans AT kotamizumoto targetingendogenousproteinsforspatialandtemporalknockdownusingauxininducibledegronincaenorhabditiselegans |