Knockout rats via embryo microinjection of zinc-finger nucleases.
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
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.