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Genetics

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JoVE Journal Genetics
CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
 

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

Article DOI: 10.3791/56844-v • 08:22 min • March 12th, 2018
March 12th, 2018 •
1,2, 1,2, 2,3,4, 1, 3, 1
1Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 2College of Life Sciences, University of Chinese Academy of Sciences, 3School of Life Science and Technology, Shanghai Tech University, 4Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences

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The clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9) system provides a promising tool for genetic engineering, and opens up the possibility of targeted integration of transgenes. We describe a homology-mediated end joining (HMEJ)-based strategy for efficient DNA targeted integration in vivo and targeted gene therapies using CRISPR/Cas9.

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CRISPR/Cas9-mediated Targeted Integration In Vivo Homology-mediated End Joining-based Strategy Genetic Tool Genetically Modified Animal Models Targeted Gene Therapies Efficiency Precise Targeted Integration Transgenes Gene Mutations High Efficiency Therapeutic Potential Xing Wang Graduate Student Protocol N2a Cells Cas9 SgRNA EGFP Expression Vectors Fluorescence Activated Cell Sorting Or FACS Lysis Buffer Proteinase K Nested PCR Outer Primers SgRNA Target Site
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