Transgenic mice for in vivo epigenome editing with CRISPR-based systems
Transgenic mice for in vivo epigenome editing with CRISPR-based systems
Abstract The discovery, characterization, and adaptation of the RNA-guided clustered regularly interspersed short palindromic repeat (CRISPR)-Cas9 system has greatly increased the ease with which genome and epigenome editing can be performed. Fusion of chromatin-modifying domains to the nuclease-deactivated form of Cas9 (dCas9) has enabled targeted gene activation or repression in both cultured cells and in vivo in animal models. However, delivery of the large dCas9 fusion proteins to target cell types and tissues is an obstacle to widespread adoption of these tools for in vivo studies. Here we describe the generation and validation of two conditional transgenic mouse lines for targeted gene regulation, Rosa26:LSL-dCas9-p300 for gene activation and Rosa26:LSL-dCas9-KRAB for gene repression. Using the dCas9p300 and dCas9KRAB transgenic mice we demonstrate activation or repression of genes in both the brain and liver in vivo, and T cells and fibroblasts ex vivo. We show gene regulation and targeted epigenetic modification with gRNAs targeting either transcriptional start sites (TSS) or distal enhancer elements, as well as corresponding changes to downstream phenotypes. These mouse lines are convenient and valuable tools for facile, temporally controlled, and tissue-restricted epigenome editing and manipulation of gene expression in vivo.
Gemberling Matthew、Rouse Douglas C.、West Anne E.、Gersbach Charles A.、Rodriguez Erica、Barrera Alejandro、Liu Fang、Cigliola Valentina、Williams Courtney、Bartelt Luke C.、Daniels Heather、Hilton Isaac B.、Ciofani Maria、Madigan Victoria J.、Bodle Josephine、Asokan Aravind、Siklenka Keith、Kantor Ariel、Poss Kenneth D.、Hazlett Mariah F.、Tonn-Eisinger Katherine R.、Li Liqing、Reddy Timothy E.
Department of Biomedical Engineering, Duke University||Center for Advanced Genomic Technologies, Duke UniversityDivision of Laboratory Animal Resources, Duke University School of MedicineCenter for Advanced Genomic Technologies, Duke University||Department of Neurobiology, Duke University School of MedicineDepartment of Biomedical Engineering, Duke University||Center for Advanced Genomic Technologies, Duke University||Department of Surgery, Duke University Medical CenterDepartment of Neurobiology, Duke University School of MedicineDepartment of Biostatistics and Bioinformatics, Duke University Medical CenterDepartment of Neurobiology, Duke University School of MedicineDepartment of Cell Biology, Duke University Medical Center||Regeneration Next, Duke UniversityCenter for Advanced Genomic Technologies, Duke University||Department of Biostatistics and Bioinformatics, Duke University Medical CenterDepartment of Biomedical Engineering, Duke University||Center for Advanced Genomic Technologies, Duke UniversityDepartment of Biomedical Engineering, Duke University||Department of Bioengineering, Rice University||Department of Biosciences, Rice UniversityCenter for Advanced Genomic Technologies, Duke University||Department of Immunology, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDepartment of Biomedical Engineering, Duke University||Center for Advanced Genomic Technologies, Duke UniversityDepartment of Biomedical Engineering, Duke University||Center for Advanced Genomic Technologies, Duke University||Department of Surgery, Duke University Medical Center||Department of Molecular Genetics and Microbiology, Duke University Medical CenterDepartment of Biostatistics and Bioinformatics, Duke University Medical CenterDepartment of Biomedical Engineering, Duke University||Center for Advanced Genomic Technologies, Duke UniversityCenter for Advanced Genomic Technologies, Duke University||Department of Cell Biology, Duke University Medical Center||Regeneration Next, Duke UniversityDepartment of Neurobiology, Duke University School of MedicineDepartment of Neurobiology, Duke University School of MedicineCenter for Advanced Genomic Technologies, Duke University||Department of Biostatistics and Bioinformatics, Duke University Medical Center
生物科学研究方法、生物科学研究技术遗传学分子生物学
Gemberling Matthew,Rouse Douglas C.,West Anne E.,Gersbach Charles A.,Rodriguez Erica,Barrera Alejandro,Liu Fang,Cigliola Valentina,Williams Courtney,Bartelt Luke C.,Daniels Heather,Hilton Isaac B.,Ciofani Maria,Madigan Victoria J.,Bodle Josephine,Asokan Aravind,Siklenka Keith,Kantor Ariel,Poss Kenneth D.,Hazlett Mariah F.,Tonn-Eisinger Katherine R.,Li Liqing,Reddy Timothy E..Transgenic mice for in vivo epigenome editing with CRISPR-based systems[EB/OL].(2025-03-28)[2025-05-29].https://www.biorxiv.org/content/10.1101/2021.03.08.434430.点此复制
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