Generation and characterization of inducible KRAB-dCas9 iPSCs from primates for cross-species CRISPRi
Fiona C Edenhofer, Anita Térmeg, Mari Ohnuki, Jessica Jocher, Zane Kliesmete, Eva Briem, Ines Hellmann, Wolfgang Enard
Abstract
Comparisons of molecular phenotypes across primates provide unique information to understand human biology and evolution, and single-cell RNA-seq CRISPR interference (CRISPRi) 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 at 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 CRISPRi screen. Hence, we provide valuable resources for performing and further extending CRISPRi in human and non-human primates.