A protocol for CRISPR-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells

Human endogenous retroviruses (HERVs) comprise many regulatory elements and can regulate host gene activity at different expression levels via multiple mechanisms. Here, we introduce a step-by-step protocol to activate or repress transcription of HERV-K(HML-2) elements using the CRISPRa and CRISPRi...

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Detalles Bibliográficos
Autores principales: Nair, Vidya Padmanabhan, Mayer, Jens, Vincendeau, Michelle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9026570/
https://www.ncbi.nlm.nih.gov/pubmed/35463468
http://dx.doi.org/10.1016/j.xpro.2022.101281
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author Nair, Vidya Padmanabhan
Mayer, Jens
Vincendeau, Michelle
author_facet Nair, Vidya Padmanabhan
Mayer, Jens
Vincendeau, Michelle
author_sort Nair, Vidya Padmanabhan
collection PubMed
description Human endogenous retroviruses (HERVs) comprise many regulatory elements and can regulate host gene activity at different expression levels via multiple mechanisms. Here, we introduce a step-by-step protocol to activate or repress transcription of HERV-K(HML-2) elements using the CRISPRa and CRISPRi technologies in human embryonic stem cells. This protocol can help deciphering the functional role of HERV-K(HML-2) elements in critical biological processes. The protocol may easily be adapted to other cell lines and HERV groups with relatively low sequence heterogeneity. For complete details on the use and execution of this protocol, please refer to Padmanabhan Nair et al. (2021).
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spelling pubmed-90265702022-04-23 A protocol for CRISPR-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells Nair, Vidya Padmanabhan Mayer, Jens Vincendeau, Michelle STAR Protoc Protocol Human endogenous retroviruses (HERVs) comprise many regulatory elements and can regulate host gene activity at different expression levels via multiple mechanisms. Here, we introduce a step-by-step protocol to activate or repress transcription of HERV-K(HML-2) elements using the CRISPRa and CRISPRi technologies in human embryonic stem cells. This protocol can help deciphering the functional role of HERV-K(HML-2) elements in critical biological processes. The protocol may easily be adapted to other cell lines and HERV groups with relatively low sequence heterogeneity. For complete details on the use and execution of this protocol, please refer to Padmanabhan Nair et al. (2021). Elsevier 2022-04-12 /pmc/articles/PMC9026570/ /pubmed/35463468 http://dx.doi.org/10.1016/j.xpro.2022.101281 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
Nair, Vidya Padmanabhan
Mayer, Jens
Vincendeau, Michelle
A protocol for CRISPR-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells
title A protocol for CRISPR-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells
title_full A protocol for CRISPR-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells
title_fullStr A protocol for CRISPR-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells
title_full_unstemmed A protocol for CRISPR-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells
title_short A protocol for CRISPR-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells
title_sort protocol for crispr-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9026570/
https://www.ncbi.nlm.nih.gov/pubmed/35463468
http://dx.doi.org/10.1016/j.xpro.2022.101281
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