Knockout rats via embryo microinjection of zinc-finger nucleases

AM Geurts, GJ Cost, Y Freyvert, B Zeitler, JC Miller… - Science, 2009 - science.org
AM Geurts, GJ Cost, Y Freyvert, B Zeitler, JC Miller, VM Choi, SS Jenkins, A Wood, X Cui…
Science, 2009science.org
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 …
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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