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Efficient bi-allelic tagging in human induced pluripotent stem cells using CRISPR.

ABSTRACT: Allelic tagging of endogenous genes enables studying gene function and transcriptional control in the native genomic context. Here, we present an efficient protocol for bi-allelic tagging of protein-coding genes with fluorescent reporters in human iPSCs using the CRISPR-Cas9-mediated homology-directed repair. We detail steps for design, cloning, electroporation, and single-cell clone isolation and validation. The tagging strategy described in this protocol is readily applicable for knockin of other reporters in diverse cell types for biomedical research.

SUBMITTER: Ren X

PROVIDER: S-EPMC9918420 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature

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Efficient bi-allelic tagging in human induced pluripotent stem cells using CRISPR.

Ren Xingjie X Takagi Maya Asami MA Shen Yin Y

STAR protocols 20230201 1

Allelic tagging of endogenous genes enables studying gene function and transcriptional control in the native genomic context. Here, we present an efficient protocol for bi-allelic tagging of protein-coding genes with fluorescent reporters in human iPSCs using the CRISPR-Cas9-mediated homology-directed repair. We detail steps for design, cloning, electroporation, and single-cell clone isolation and validation. The tagging strategy described in this protocol is readily applicable for knockin of ot ...[more]

PMID: 36853689

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