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Knockout Rats via Embryo Microinjection of Zinc-Finger Nucleases

Aron M. Geurts,Gregory J. Cost,Yevgeniy Freyvert,Bryan Zeitler,Jeffrey C. Miller,Vivian M. Choi,Shirin S Jenkins,Adam Wood,Xiaoxia Cui,Xiangdong Meng,Anna I Vincent,Stephen Lam,Mieczyslaw Michalkiewicz,Rebecca Schilling,Jamie Foeckler,Shawn Kalloway,Hartmut Weiler,Séverine Menoret,Ignacio Anegón,Gregory D. Davis,Lei Zhang,Edward J. Rebar,Philip D. Gregory,Fyodor D. Urnov,Howard J. Jacob,Roland Buelow

Zinc finger nuclease

Zinc finger

Biology

2009

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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“Knockout Rats via Embryo Microinjection of Zinc-Finger Nucleases” is a paper by Aron M. Geurts Gregory J. Cost Yevgeniy Freyvert Bryan Zeitler Jeffrey C. Miller Vivian M. Choi Shirin S Jenkins Adam Wood Xiaoxia Cui Xiangdong Meng Anna I Vincent Stephen Lam Mieczyslaw Michalkiewicz Rebecca Schilling Jamie Foeckler Shawn Kalloway Hartmut Weiler Séverine Menoret Ignacio Anegón Gregory D. Davis Lei Zhang Edward J. Rebar Philip D. Gregory Fyodor D. Urnov Howard J. Jacob Roland Buelow published in 2009. It has an Open Access status of “green”. You can read and download a PDF Full Text of this paper here.