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Generation and characterization of inducible KRAB-dCas9 iPSCs from primates for cross-species CRISPRi

ABSTRACT: Comparisons of molecular phenotypes across primates provide unique information to understand human biology and evolution and single-cell RNA-seq CRISPR interference screens are a powerful approach to analyze them. Here, we generate and validate three human, three gorilla and two cynomolgus iPS cell lines that carry a dox-inducible KRAB-dCas9 construct in the AAVS1 locus. We show that despite variable expression levels of KRAB-dCas9 among lines, comparable downregulation of target genes and comparable phenotypic effects are observed in a single-cell RNA-seq CRISPR interference screen. Hence, we provide valuable resources for performing and further extending CRISPRi screens in human and non-human primates.

ORGANISM(S): Homo sapiens Gorilla gorilla Macaca fascicularis

PROVIDER: GSE255980 | GEO | 2024/02/20

REPOSITORIES: GEO

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Generation and characterization of inducible KRAB-dCas9 iPSCs from primates for cross-species CRISPRi

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