show Abstracthide AbstractComparisons 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. Overall design: Two human and one cynomolgus KRAB-dCas9 iPS cell line were transduced with species-specific lentiviral gRNA libraries. Using the 10X Genomics platform, gene-expression and CRISPR libraries for capturing the gRNAs were generated and sequenced.