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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Efficient targeted gene disruption in Xenopus embryos using engineered transcription activator-like effector nucleases (TALENs)
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Efficient targeted gene disruption in Xenopus embryos using engineered transcription activator-like effector nucleases (TALENs)

机译：使用工程转录激活因子样效应子核酸酶（TALEN）在非洲爪蟾胚胎中有效靶向基因破坏。

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摘要

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.
机译：转录激活子样效应核酸酶（TALENs）是一种定向基因破坏的方法，并已被证明在各种动物模型中都是有效的。在这里，我们报告TalENs可以可靠地高效地诱导非洲爪蟾胚胎中的体细胞突变，并且这种突变通过种系传递是可遗传的。我们修改了TalEN组装的Golden Gate方法，使该产品适合RNA转录和显微注射到非洲爪蟾胚胎中。构建了八对TalEN，以靶向八个非洲爪蟾基因，所有这些都导致在目标基因座处的插入缺失突变，效率高达95.7％。此外，由TalENs诱导的突变已通过种系高效传递至F_1青蛙。结合简单可靠的基于PCR的方法检测TalEN引起的突变，我们的结果表明TalENs是非洲爪蟾中靶向基因编辑/敲除的有效工具。

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  • 作者

    Yong Lei; Xiaogang Guo; Yun Liu; Yang Cao; Yi Deng; Xiongfeng Chen; Christopher H. K. Cheng; Igor B. Dawid; Yonglong Chen; Hui Zhao;

  • 作者单位

    School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, People's Republic of China;

    Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, People's Republic of CHina;

    School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, People's Republic of China;

    Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, People's Republic of CHina;

    Department of Biology, South University of Science and Technology of China, Shenzhen 518055, People's Republic of China;

    Advanced Biomedical Computing Center, National Cancer Institute, National Institutes of Health, Frederick, MD 21702;

    School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, People's Republic of China,Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen 518057, People's Republic of China;

    Program in Genomics of Differentiation, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892;

    Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, People's Republic of CHina;

    School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, People's Republic of China,Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen 518057, People's Republic of China;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
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  • 关键词

    genome editing; heritable mutagenesis; mutagenesis detection reverse genetics; genome engineering;

    机译：基因组编辑;遗传诱变;诱变检测反向遗传学;基因组工程;

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