CRISPR interference provides increased cell type-specificity compared to the Cre-loxP system

Cre-mediated recombination is frequently used for cell type-specific loss of function (LOF) studies. A major limitation of this system is recombination in unwanted cell types. CRISPR interference (CRISPRi) has been used effectively for global LOF in mice. However, cell type-specific CRISPRi, independent of recombination-based systems, has not been reported. To test the feasibility of cell type-specific CRISPRi, we produced two novel knock-in mouse models that achieve gene suppression when used together: one expressing dCas9::KRAB under the control of a cell type-specific promoter and the other expressing a single guide RNA from a safe harbor locus. We then compared the phenotypes of mice in which the same gene was targeted by either CRISPRi or the Cre-loxP system, with cell specificity conferred by Dmp1 regulatory elements in both cases. We demonstrate that CRISPRi is effective for cell type-specific LOF and that it provides improved cell type-specificity compared to the Cre-loxP system. [Display omitted] •In vivo global gene knockdown by CRISPRi can be as effective as gene knockout•CRISPRi can be adapted to perform in vivo cell type-specific loss of function•Cell type-specific loss of function using CRISPRi is more specific than Cre-loxP•Knockdown by cell type-specific CRISPRi persists up to 12 months of age Genetics; Molecular Genetics