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Science 24 July 2009:
Vol. 325. no. 5939, p. 433
DOI: 10.1126/science.1172447
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Brevia

Knockout Rats via Embryo Microinjection of Zinc-Finger Nucleases

Aron M. Geurts,1,2,* Gregory J. Cost,3,* Yevgeniy Freyvert,3 Bryan Zeitler,3 Jeffrey C. Miller,3 Vivian M. Choi,3 Shirin S. Jenkins,3 Adam Wood,4 Xiaoxia Cui,4 Xiangdong Meng,3 Anna Vincent,3 Stephen Lam,3 Mieczyslaw Michalkiewicz,1,2 Rebecca Schilling,1,2 Jamie Foeckler,3 Shawn Kalloway,3 Hartmut Weiler,1,2 Séverine Ménoret,5 Ignacio Anegon,5 Gregory D. Davis,4 Lei Zhang,3 Edward J. Rebar,3 Philip D. Gregory,3 Fyodor D. Urnov,3 Howard J. Jacob,1,2,6,{dagger} Roland Buelow7,{dagger}

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.

1 Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, WI 52336, USA.
2 Department of Physiology, Medical College of Wisconsin, Milwaukee, WI 52336, USA.
3 Sangamo BioSciences, Incorporated, Richmond, CA 94804, USA.
4 Sigma-Aldrich Biotechnology, St. Louis, MO 63103, USA.
5 INSERM, UMR 643, CHU, Nantes, Université de Nantes, 44322 Nantes, France.
6 Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI 52336, USA.
7 Open Monoclonal Technology, Incorporated, Palo Alto, CA 94303, USA.
{dagger} To whom correspondence should be addressed. E-mail: jacob{at}mcw.edu (H.J.J.); rbuelow{at}omtinc.net (R.B.)

* These authors contributed equally to this work.

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THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
"Gene Targeting in the Rat? Cut it out!".
B. M. G. Smits and M. N. Gould (2009)
Mol. Interv. 9, 226-229
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