Generation of locus-specific degradable tag knock-ins in mouse and human cell lines
© 2021 The Author(s)..
Protein degradation technologies represent a powerful functional genomics tool, allowing fast and controllable target protein depletion. Establishing these systems requires a knock-in of the degradation tag into both endogenous target gene alleles. Here, we provide a step-by-step protocol for the efficient generation of biallelic degradation tag knock-ins in mouse and human cell lines using CRISPR-Cas9. We use knockin of an endogenous Kansl3 degradation tag in mouse embryonic stem (ES) cells as an example but provide modifications for application in other cell types. For complete details on the use and execution of this protocol, please refer to Radzisheuskaya et al. (2021).
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:2 |
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| Enthalten in: |
STAR protocols - 2(2021), 2 vom: 18. Juni, Seite 100575 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Damhofer, Helene [VerfasserIn] |
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| Links: |
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| Themen: |
CRISPR |
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| Anmerkungen: |
Date Completed 21.03.2022 Date Revised 21.03.2022 published: Electronic-eCollection Citation Status MEDLINE |
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| doi: |
10.1016/j.xpro.2021.100575 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM326987487 |
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| 500 | |a Date Revised 21.03.2022 | ||
| 500 | |a published: Electronic-eCollection | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2021 The Author(s). | ||
| 520 | |a Protein degradation technologies represent a powerful functional genomics tool, allowing fast and controllable target protein depletion. Establishing these systems requires a knock-in of the degradation tag into both endogenous target gene alleles. Here, we provide a step-by-step protocol for the efficient generation of biallelic degradation tag knock-ins in mouse and human cell lines using CRISPR-Cas9. We use knockin of an endogenous Kansl3 degradation tag in mouse embryonic stem (ES) cells as an example but provide modifications for application in other cell types. For complete details on the use and execution of this protocol, please refer to Radzisheuskaya et al. (2021) | ||
| 650 | 4 | |a Journal Article | |
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| 650 | 4 | |a Genetics | |
| 650 | 4 | |a Molecular Biology | |
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| 700 | 1 | |a Helin, Kristian |e verfasserin |4 aut | |
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