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CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
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
Chapters
- 00:04Title
- 00:54sgRNA Surveyor Nuclease Assay
- 02:50Cas9 mRNA Production
- 04:11sgRNA Production
- 05:30Embryo Collection, Microinjection, and In Vitro Culture
- 06:43Results: Superior Integration with Cas9
- 07:46Conclusion
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.
