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Optimized protocol for gene editing in adipocytes using CRISPR-Cas9 technology

We present a detailed protocol for gene editing in adipocytes using the CRISPR-Cas technology. This protocol describes sgRNA design, preparation of lentiCRISPR-sgRNA vectors, functional validation of sgRNAs, preparation of lentiviruses, and lentiviruses transduction in adipocytes. Moreover, an optim...

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Detalles Bibliográficos
Autores principales: Qiu, Yan, Ding, Qiurong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7844572/
https://www.ncbi.nlm.nih.gov/pubmed/33554142
http://dx.doi.org/10.1016/j.xpro.2021.100307
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author Qiu, Yan
Ding, Qiurong
author_facet Qiu, Yan
Ding, Qiurong
author_sort Qiu, Yan
collection PubMed
description We present a detailed protocol for gene editing in adipocytes using the CRISPR-Cas technology. This protocol describes sgRNA design, preparation of lentiCRISPR-sgRNA vectors, functional validation of sgRNAs, preparation of lentiviruses, and lentiviruses transduction in adipocytes. Moreover, an optimized method of gene editing using the lentiCRISPRv2 vector expressing two sgRNAs targeting two different genes has also been described. For complete details on the use and execution of this protocol, please refer to Qiu et al. (2020).
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spelling pubmed-78445722021-02-04 Optimized protocol for gene editing in adipocytes using CRISPR-Cas9 technology Qiu, Yan Ding, Qiurong STAR Protoc Protocol We present a detailed protocol for gene editing in adipocytes using the CRISPR-Cas technology. This protocol describes sgRNA design, preparation of lentiCRISPR-sgRNA vectors, functional validation of sgRNAs, preparation of lentiviruses, and lentiviruses transduction in adipocytes. Moreover, an optimized method of gene editing using the lentiCRISPRv2 vector expressing two sgRNAs targeting two different genes has also been described. For complete details on the use and execution of this protocol, please refer to Qiu et al. (2020). Elsevier 2021-01-27 /pmc/articles/PMC7844572/ /pubmed/33554142 http://dx.doi.org/10.1016/j.xpro.2021.100307 Text en © 2021 The Author(s) http://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
Qiu, Yan
Ding, Qiurong
Optimized protocol for gene editing in adipocytes using CRISPR-Cas9 technology
title Optimized protocol for gene editing in adipocytes using CRISPR-Cas9 technology
title_full Optimized protocol for gene editing in adipocytes using CRISPR-Cas9 technology
title_fullStr Optimized protocol for gene editing in adipocytes using CRISPR-Cas9 technology
title_full_unstemmed Optimized protocol for gene editing in adipocytes using CRISPR-Cas9 technology
title_short Optimized protocol for gene editing in adipocytes using CRISPR-Cas9 technology
title_sort optimized protocol for gene editing in adipocytes using crispr-cas9 technology
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7844572/
https://www.ncbi.nlm.nih.gov/pubmed/33554142
http://dx.doi.org/10.1016/j.xpro.2021.100307
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