High-efficiency and heritable gene targeting in mouse by transcription activator-like effector nucleases.
High-efficiency and heritable gene targeting in mouse by transcription activator-like effector nucleases.
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通过类转录激活因子效应核酸酶在小鼠中实现高效、可遗传的基因靶向
DOI：
10.1093/nar/gkt258
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发表时间：
2013-06
影响因子：
14.9
通讯作者：
Li D
中科院分区：
文献类型：
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作者：
Qiu Z;Liu M;Chen Z;Shao Y;Pan H;Wei G;Yu C;Zhang L;Li X;Wang P;Fan HY;Du B;Liu B;Liu M;Li D
转录激活子样效应核酸酶 (TALEN) 是一种在各种动物模型中靶向基因破坏的强大新方法，但人们对它们在广泛使用的哺乳动物模型生物小家鼠中的活性知之甚少。在这里，我们报告将体外转录的TALEN对的信使RNA直接注射到小鼠受精卵中诱导体细胞突变，并通过种系传递稳定地传递给下一代。通过针对10个目标基因中的每一个构建一对TALEN，获得每个基因的突变F0小鼠，突变率范围为13%至67%，平均为健康新生儿总数的约40%，C57BL/6和FVB/N遗传背景之间没有显着差异。一对 TALEN 与每一侧的预期目标序列存在单一错配，未能产生任何突变。此外，获得了高效的种系传递，因为所有测试的 F0 创始人都将突变传递给了 F1 小鼠。此外，我们还观察到Lepr基因的一个双等位基因突变体（编码瘦素受体）具有与db/db小鼠相似的糖尿病表型。总之，我们的结果表明，TALEN 是一种有效的遗传工具，可以在具有不同遗传背景的小鼠中高效且具有遗传性地快速进行基因破坏。
Transcription activator-like effector nucleases (TALENs) are a powerful new approach for targeted gene disruption in various animal models, but little is known about their activities in Mus musculus, the widely used mammalian model organism. Here, we report that direct injection of in vitro transcribed messenger RNA of TALEN pairs into mouse zygotes induced somatic mutations, which were stably passed to the next generation through germ-line transmission. With one TALEN pair constructed for each of 10 target genes, mutant F0 mice for each gene were obtained with the mutation rate ranged from 13 to 67% and an average of ∼40% of total healthy newborns with no significant differences between C57BL/6 and FVB/N genetic background. One TALEN pair with single mismatch to their intended target sequence in each side failed to yield any mutation. Furthermore, highly efficient germ-line transmission was obtained, as all the F0 founders tested transmitted the mutations to F1 mice. In addition, we also observed that one bi-allele mutant founder of Lepr gene, encoding Leptin receptor, had similar diabetic phenotype as db/db mouse. Together, our results suggest that TALENs are an effective genetic tool for rapid gene disruption with high efficiency and heritability in mouse with distinct genetic background.
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影响因子：
14.9
作者：
Li T;Huang S;Jiang WZ;Wright D;Spalding MH;Weeks DP;Yang B
通讯作者：
Yang B
影响因子：
64.8
作者：
MANSOUR, SL;THOMAS, KR;CAPECCHI, MR
通讯作者：
CAPECCHI, MR
影响因子：
14.9
作者：
Cade L;Reyon D;Hwang WY;Tsai SQ;Patel S;Khayter C;Joung JK;Sander JD;Peterson RT;Yeh JR
通讯作者：
Yeh JR
影响因子：
56.9
作者：
Boch, Jens;Scholze, Heidi;Bonas, Ulla
通讯作者：
Bonas, Ulla
影响因子：
16.8
作者：
通讯作者：
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