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

Edenhofer, Fiona C.; Térmeg, Anita; Ohnuki, Mari; Jocher, Jessica; Kliesmete, Zane; Briem, Eva; Hellmann, Ines; Enard, Wolfgang (2024): Generation and characterization of inducible KRAB-dCas9 iPSCs from primates for cross-species CRISPRi. iScience, 27 (6). p. 110090. ISSN 25890042

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.

• Doc-Type: Article (LMU)
• Organisational unit (Faculties): 19 Biology
• DFG subject classification of scientific disciplines: Life sciences
• DFG: Funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - 458888224
• DFG: Funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - 458247426
• DFG: Funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - 491502892