Modeling disease mutations by gene targeting in one-cell mouse embryos
Modeling disease mutations by gene targeting in one-cell mouse embryos
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DOI：
10.1073/pnas.1121203109
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发表时间：
2012-06-12
影响因子：
11.1
通讯作者：
Kuehn, Ralf
中科院分区：
文献类型：
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作者：
Meyer, Melanie;Ortiz, Oskar;Kuehn, Ralf
单细胞胚胎中锌指核酸酶的基因靶向为小鼠、大鼠和兔子提供了一种快速诱变方法。该技术最近已被用于通过核苷酸的删除或插入来创建敲除和敲入突变体。在这里，我们在单细胞小鼠胚胎中应用锌指核酸酶来产生带有单核苷酸或密码子替代的疾病相关突变体。使用基因靶向载体或合成的寡脱氧核苷酸作为同源重组的模板，我们将错义和沉默突变引入 Rab38 基因，编码调节细胞内囊泡运输的小 GTPase。这些结果证明了在单细胞胚胎中进行无缝基因编辑以创建遗传疾病模型并建立合成寡脱氧核苷酸作为简化诱变工具的可行性。
Gene targeting by zinc-finger nucleases in one-cell embryos provides an expedite mutagenesis approach in mice, rats, and rabbits. This technology has been recently used to create knockout and knockin mutants through the deletion or insertion of nucleotides. Here we apply zinc-finger nucleases in one-cell mouse embryos to generate disease-related mutants harboring single nucleotide or codon replacements. Using a gene-targeting vector or a synthetic oligodesoxynucleotide as template for homologous recombination, we introduced missense and silent mutations into the Rab38 gene, encoding a small GTPase that regulates intracellular vesicle trafficking. These results demonstrate the feasibility of seamless gene editing in one-cell embryos to create genetic disease models and establish synthetic oligodesoxynucleotides as a simplified mutagenesis tool.