School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, People's Republic of China.
Proc Natl Acad Sci U S A. 2012 Oct 23;109(43):17484-9. doi: 10.1073/pnas.1215421109. Epub 2012 Oct 8.
Transcription activator-like effector nucleases (TALENs) are an approach for directed gene disruption and have been proved to be effective in various animal models. Here, we report that TALENs can induce somatic mutations in Xenopus embryos with reliably high efficiency and that such mutations are heritable through germ-line transmission. We modified the Golden Gate method for TALEN assembly to make the product suitable for RNA transcription and microinjection into Xenopus embryos. Eight pairs of TALENs were constructed to target eight Xenopus genes, and all resulted in indel mutations with high efficiencies of up to 95.7% at the targeted loci. Furthermore, mutations induced by TALENs were highly efficiently passed through the germ line to F(1) frogs. Together with simple and reliable PCR-based approaches for detecting TALEN-induced mutations, our results indicate that TALENs are an effective tool for targeted gene editing/knockout in Xenopus.
转录激活因子样效应物核酸酶（TALENs）是一种定向基因敲除的方法，已被证明在各种动物模型中非常有效。在这里，我们报告 TALENs 可以以可靠的高效率在爪蟾胚胎中诱导体细胞突变，并且这种突变可以通过种系传递而遗传。我们修改了 Golden Gate 方法来组装 TALEN，使产物适合 RNA 转录和微注射到爪蟾胚胎中。构建了 8 对 TALEN 来靶向 8 个爪蟾基因，所有靶向位点的插入缺失突变效率高达 95.7%。此外，TALEN 诱导的突变通过种系高效传递到 F1 青蛙。结合基于 PCR 的简单可靠方法检测 TALEN 诱导的突变，我们的结果表明 TALENs 是一种在爪蟾中进行靶向基因编辑/敲除的有效工具。
