CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development

This protocol describes the design and use of CRISPRi-mediated transcriptional silencing in human iPSCs, for loss-of-function studies in brain development research. The protocol avoids single cell selection, thereby eliminating side effects of clonal expansion and sites of viral integration. We also...

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Detalles Bibliográficos
Autores principales: Johansson, Pia Annette, Adami, Anita, Jakobsson, Johan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9038555/
https://www.ncbi.nlm.nih.gov/pubmed/35496780
http://dx.doi.org/10.1016/j.xpro.2022.101285
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author Johansson, Pia Annette
Adami, Anita
Jakobsson, Johan
author_facet Johansson, Pia Annette
Adami, Anita
Jakobsson, Johan
author_sort Johansson, Pia Annette
collection PubMed
description This protocol describes the design and use of CRISPRi-mediated transcriptional silencing in human iPSCs, for loss-of-function studies in brain development research. The protocol avoids single cell selection, thereby eliminating side effects of clonal expansion and sites of viral integration. We also describe a neural progenitor differentiation protocol and discuss the challenges of target-specific lentiviral silencing, efficient silencing levels, and off-target effects. For complete details on the use and execution of this protocol, please refer to Johansson et al. (2022).
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spelling pubmed-90385552022-04-27 CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development Johansson, Pia Annette Adami, Anita Jakobsson, Johan STAR Protoc Protocol This protocol describes the design and use of CRISPRi-mediated transcriptional silencing in human iPSCs, for loss-of-function studies in brain development research. The protocol avoids single cell selection, thereby eliminating side effects of clonal expansion and sites of viral integration. We also describe a neural progenitor differentiation protocol and discuss the challenges of target-specific lentiviral silencing, efficient silencing levels, and off-target effects. For complete details on the use and execution of this protocol, please refer to Johansson et al. (2022). Elsevier 2022-04-13 /pmc/articles/PMC9038555/ /pubmed/35496780 http://dx.doi.org/10.1016/j.xpro.2022.101285 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Johansson, Pia Annette
Adami, Anita
Jakobsson, Johan
CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development
title CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development
title_full CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development
title_fullStr CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development
title_full_unstemmed CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development
title_short CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development
title_sort crispri-mediated transcriptional silencing in ipscs for the study of human brain development
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9038555/
https://www.ncbi.nlm.nih.gov/pubmed/35496780
http://dx.doi.org/10.1016/j.xpro.2022.101285
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