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Knockout rats via embryo microinjection of zinc-finger nucleases.

Authors: Geurts, AM  Cost, GJ  Freyvert, Y  Zeitler, B  Miller, JC  Choi, VM  Jenkins, SS  Wood, A  Cui, X  Meng, X  Vincent, A  Lam, S  Michalkiewicz, M  Schilling, R  Foeckler, J  Kalloway, S  Weiler, H  Menoret, S  Anegon, I  Davis, GD  Zhang, L  Rebar, EJ  Gregory, PD  Urnov, FD  Jacob, HJ  Buelow, R 
Citation: Geurts AM, etal., Science. 2009 Jul 24;325(5939):433.
Pubmed: (View Article at PubMed) PMID:19628861
DOI: Full-text: DOI:10.1126/science.1172447

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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RGD Object Information
RGD ID: 4131263
Created: 2010-08-19
Species: All species
Last Modified: 2010-08-19
Status: ACTIVE


RGD is funded by grant HL64541 from the National Heart, Lung, and Blood Institute on behalf of the NIH.