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

doi:10.1016/j.isci.2023.107428

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

iScience. 2023 Jul 20;26(8):107428. doi: 10.1016/j.isci.2023.107428. eCollection 2023 Aug 18.

Authors

Dominique J Laster¹, Nisreen S Akel¹, James A Hendrixson¹, Alicen James¹, Julie A Crawford^{2

3}, Qiang Fu^{2

3}, Stuart B Berryhill^{2

3}, Jeff D Thostenson⁴, Intawat Nookaew^{2

5}, Charles A O'Brien^{2

3}, Melda Onal^{1

2}

Affiliations

¹ Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
² Center for Musculoskeletal Disease Research (CMDR), University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
³ Division of Endocrinology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
⁴ Department of Biostatistics, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
⁵ Department of Biomedical Informatics, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.

Abstract

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.

Keywords: Genetics; Molecular Genetics.

Abstract

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