Department of Pediatrics, Division of Genes and Development; Department of Microbiology and Physiological Systems; Horae Gene Therapy Center
Biomedical Engineering and Bioengineering | Computational Biology | Therapeutics
Recent advances using CRISPR-Cas9 approaches have dramatically enhanced the ease for genetic manipulation in rodents. Notwithstanding, the methods to deliver nucleic acids into pre-implantation embryos have hardly changed since the original description of mouse transgenesis more than 30 years ago. Here we report a novel strategy to generate genetically modified mice by transduction of CRISPR-Cas9 components into pre-implantation mouse embryos via recombinant adeno-associated viruses (rAAVs). Using this approach, we efficiently generated a variety of targeted mutations in explanted embryos, including indel events produced by non-homologous end joining and tailored mutations using homology-directed repair. We also achieved gene modification in vivo by direct delivery of rAAV particles into the oviduct of pregnant females. Our approach greatly simplifies the generation of genetically modified mice and, more importantly, opens the door for streamlined gene editing in other mammalian species.
CRISPR-Cas9 genome editing, Genetic engineering
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Copyright © The Author(s) 2018. Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
DOI of Published Version
Nat Commun. 2018 Jan 29;9(1):412. doi: 10.1038/s41467-017-02706-7. Link to article on publisher's site
Yoon, Yeonsoo; Wang, Dan; Tai, Phillip W. L.; Riley, Joy; Gao, Guangping; and Rivera-Perez, Jaime A., "Streamlined ex vivo and in vivo genome editing in mouse embryos using recombinant adeno-associated viruses" (2018). Pediatric Publications and Presentations. 204.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.