Knockout Rats via Embryo Microinjection of Zinc-Finger Nucleases

Aron M. Geurts(Medical College of Wisconsin), Gregory J. Cost(Sangamo BioSciences (United States)), Yevgeniy Freyvert(Sangamo BioSciences (United States)), Bryan Zeitler(Sangamo BioSciences (United States)), Jeffrey C. Miller(Sangamo BioSciences (United States)), Vivian M. Choi(Sangamo BioSciences (United States)), Shirin S Jenkins(Sangamo BioSciences (United States)), Adam J. Wood(Sigma Research (United States)), Xiaoxia Cui(Sigma Research (United States)), Xiangdong Meng(Sangamo BioSciences (United States)), Anna Vincent(Sangamo BioSciences (United States)), Stephen C.-T. Lam(Sangamo BioSciences (United States)), Mieczyslaw Michalkiewicz(Medical College of Wisconsin), Rebecca Schilling(Medical College of Wisconsin), Jamie Foeckler(Sangamo BioSciences (United States)), Shawn Kalloway(Sangamo BioSciences (United States)), Hartmut Weiler(Medical College of Wisconsin), Séverine Ménoret(Inserm), Ignacio Anegón(Inserm), Gregory D. Davis(Sigma Research (United States)), Lei Zhang(Sangamo BioSciences (United States)), Edward J. Rebar(Sangamo BioSciences (United States)), Philip D. Gregory(Sangamo BioSciences (United States)), Fyodor D. Urnov(Sangamo BioSciences (United States)), Howard J. Jacob(Medical College of Wisconsin), Roland Buelow(Open Society)
Science
July 23, 2009
10.1126/science.1172447
Cited by 918Open Access
Full Text

Abstract

The toolbox of rat genetics currently lacks the ability to introduce site-directed, heritable mutations into the genome to create knockout animals. By using engineered zinc-finger nucleases (ZFNs) designed to target an integrated reporter and two endogenous rat genes, Immunoglobulin M (IgM) and Rab38, we demonstrate that a single injection of DNA or messenger RNA encoding ZFNs into the one-cell rat embryo leads to a high frequency of animals carrying 25 to 100% disruption at the target locus. These mutations are faithfully and efficiently transmitted through the germline. Our data demonstrate the feasibility of targeted gene disruption in multiple rat strains within 4 months time, paving the way to a humanized monoclonal antibody platform and additional human disease models.

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