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

ABSTRACT: 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).

SUBMITTER: Zhou Z

PROVIDER: S-EPMC8164093 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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

Zhou Zhongshu Z Ma Lin L Zhang Xiaoqing X

STAR protocols 20210520 2

<i>PAX6</i> 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 <i>PAX6</i> 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 execu ...[more]

PMID: 34095862

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