Streamlined ex vivo and in vivo genome editing in mouse embryos using recombinant adeno-associated viruses

Yeonsoo Yoon, Dan Wang, Phillip W. L. Tai, Joy Riley, Guangping Gao () and Jaime A. Rivera-Pérez ()
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Yeonsoo Yoon: University of Massachusetts Medical School
Dan Wang: University of Massachusetts Medical School
Phillip W. L. Tai: University of Massachusetts Medical School
Joy Riley: University of Massachusetts Medical School
Guangping Gao: University of Massachusetts Medical School
Jaime A. Rivera-Pérez: University of Massachusetts Medical School

Nature Communications, 2018, vol. 9, issue 1, 1-12

Abstract: Abstract 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.

Date: 2018
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DOI: 10.1038/s41467-017-02706-7

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