CRISPR interference provides increased cell type-specificity compared to the Cre-loxP system
Dominique J. Laster,
Nisreen S. Akel,
James A. Hendrixson,
Alicen James,
Julie A. Crawford,
Qiang Fu,
Stuart B. Berryhill,
Jeff D. Thostenson,
Intawat Nookaew,
Charles A. O’Brien,
Melda Onal
Affiliations
Dominique J. Laster
Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA
Nisreen S. Akel
Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA
James A. Hendrixson
Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA
Alicen James
Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA
Julie A. Crawford
Center for Musculoskeletal Disease Research (CMDR), University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA; Division of Endocrinology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA
Qiang Fu
Center for Musculoskeletal Disease Research (CMDR), University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA; Division of Endocrinology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA
Stuart B. Berryhill
Center for Musculoskeletal Disease Research (CMDR), University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA; Division of Endocrinology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA
Jeff D. Thostenson
Department of Biostatistics, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA
Intawat Nookaew
Center for Musculoskeletal Disease Research (CMDR), University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA; Department of Biomedical Informatics, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA
Charles A. O’Brien
Center for Musculoskeletal Disease Research (CMDR), University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA; Division of Endocrinology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA
Melda Onal
Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA; Center for Musculoskeletal Disease Research (CMDR), University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA; Corresponding author
Summary: 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.
