通过向胚胎显微注射转录激活样效应因子核酸酶（TALENs）来生成靶向小鼠突变体。
Generation of targeted mouse mutants by embryo microinjection of TALENs.
作者信息
Wefers Benedikt, Ortiz Oskar, Wurst Wolfgang, Kühn Ralf
机构信息
Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Munich, Germany.
Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Munich, Germany; Technische Universität München, 85350 Freising-Weihenstephan, Germany; Deutsches Zentrum für Neurodegenerative Erkrankungen e.V. (DZNE), 80336 Munich, Germany; Max Planck Institute of Psychiatry, 80804 Munich, Germany.
出版信息
Methods. 2014 Aug 15;69(1):94-101. doi: 10.1016/j.ymeth.2014.01.002. Epub 2014 Jan 11.
Gene engineering for generating targeted mouse mutants is a key technology for biomedical research. Using TALENs as nucleases to induce targeted double-strand breaks, the mouse genome can be directly modified in zygotes in a single step, without the need for embryonic stem cells. Thereby, knockout and knockin alleles can be generated fast and efficiently by embryo microinjection of TALEN mRNAs and targeting vectors. In this article we present an introduction into the TALEN technology and provide protocols for the application of TALENs in mouse zygotes.
生成靶向小鼠突变体的基因工程是生物医学研究的一项关键技术。使用转录激活样效应因子核酸酶（TALENs）作为核酸酶来诱导靶向双链断裂，无需胚胎干细胞，即可在一步操作中直接对受精卵中的小鼠基因组进行修饰。因此，通过向胚胎显微注射TALEN mRNA和靶向载体，能够快速高效地产生基因敲除和基因敲入等位基因。在本文中，我们介绍了TALEN技术，并提供了在小鼠受精卵中应用TALENs的实验方案。
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