Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination

PAX6 is a key determinant of human neuroectoderm cell fate. Here, we describe a protocol for genome-scale CRISPR screening for use in genetically engineered human pluripotent stem cells (hPSCs). Using the germ layer reporter PAX6 and an inducible CRISPR/Cas9 knockout system, we describe how to ident...

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Detalles Bibliográficos
Autores principales: Zhou, Zhongshu, Ma, Lin, Zhang, Xiaoqing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8164093/
https://www.ncbi.nlm.nih.gov/pubmed/34095862
http://dx.doi.org/10.1016/j.xpro.2021.100548
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author Zhou, Zhongshu
Ma, Lin
Zhang, Xiaoqing
author_facet Zhou, Zhongshu
Ma, Lin
Zhang, Xiaoqing
author_sort Zhou, Zhongshu
collection PubMed
description PAX6 is a key determinant of human neuroectoderm cell fate. Here, we describe a protocol for genome-scale CRISPR screening for use in genetically engineered human pluripotent stem cells (hPSCs). Using the germ layer reporter PAX6 and an inducible CRISPR/Cas9 knockout system, we describe how to identify lineage-specific preventing genes. This protocol can be applied for use with other reporter genes to study cell fate determination in hPSCs. For complete details on the use and execution of this protocol, please refer to Xu et al. (2021).
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spelling pubmed-81640932021-06-04 Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination Zhou, Zhongshu Ma, Lin Zhang, Xiaoqing STAR Protoc Protocol PAX6 is a key determinant of human neuroectoderm cell fate. Here, we describe a protocol for genome-scale CRISPR screening for use in genetically engineered human pluripotent stem cells (hPSCs). Using the germ layer reporter PAX6 and an inducible CRISPR/Cas9 knockout system, we describe how to identify lineage-specific preventing genes. This protocol can be applied for use with other reporter genes to study cell fate determination in hPSCs. For complete details on the use and execution of this protocol, please refer to Xu et al. (2021). Elsevier 2021-05-20 /pmc/articles/PMC8164093/ /pubmed/34095862 http://dx.doi.org/10.1016/j.xpro.2021.100548 Text en © 2021 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
Zhou, Zhongshu
Ma, Lin
Zhang, Xiaoqing
Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination
title Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination
title_full Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination
title_fullStr Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination
title_full_unstemmed Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination
title_short Protocol for genome-scale CRISPR screening in engineered lineage reporter hPSCs to study cell fate determination
title_sort protocol for genome-scale crispr screening in engineered lineage reporter hpscs to study cell fate determination
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8164093/
https://www.ncbi.nlm.nih.gov/pubmed/34095862
http://dx.doi.org/10.1016/j.xpro.2021.100548
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