Geurts Aron M, Cost Gregory J, Freyvert Yevgeniy, Zeitler Bryan, Miller Jeffrey C, Choi Vivian M, Jenkins Shirin S, Wood Adam, Cui Xiaoxia, Meng Xiangdong, Vincent Anna, Lam Stephen, Michalkiewicz Mieczyslaw, Schilling Rebecca, Foeckler Jamie, Kalloway Shawn, Weiler Hartmut, Ménoret Séverine, Anegon Ignacio, Davis Gregory D, Zhang Lei, Rebar Edward J, Gregory Philip D, Urnov Fyodor D, Jacob Howard J, Buelow Roland
Human and Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, WI 52336, USA.
Science. 2009 Jul 24;325(5939):433. 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.
目前，大鼠遗传学的工具库缺乏将位点定向的可遗传突变引入基因组以创建基因敲除动物的能力。通过使用设计用于靶向整合报告基因以及两个内源性大鼠基因（免疫球蛋白M（IgM）和Rab38）的工程化锌指核酸酶（ZFN），我们证明向单细胞大鼠胚胎单次注射编码ZFN的DNA或信使RNA会导致在目标位点携带25%至100%破坏的动物出现高频。这些突变通过种系被忠实地、高效地传递。我们的数据证明了在4个月时间内在多个大鼠品系中进行靶向基因破坏的可行性，为人类化单克隆抗体平台和其他人类疾病模型铺平了道路。
