A highly effective TALEN-mediated approach for targeted gene disruption in Xenopus tropicalis and zebrafish
A highly effective TALEN-mediated approach for targeted gene disruption in Xenopus tropicalis and zebrafish
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一种高效的 TALEN 介导方法，用于热带爪蟾和斑马鱼的靶向基因破坏
DOI：
10.1016/j.ymeth.2014.02.011
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发表时间：
2014-08-15
期刊：
影响因子：
4.8
通讯作者：
Cheng, Christopher H. K.
中科院分区：
文献类型：
--
作者：
Liu, Yun;Luo, Daji;Cheng, Christopher H. K.
转录激活物样效应核酸酶(TALENS)是一种很有前途的干扰预期基因组座位的方法。高效TALEN的组装对于成功的基因组编辑至关重要。最近，我们报道了一个方便而强大的平台来构建定制的TALEN。该平台产生的TALEN已被证明对非洲爪哇和斑马鱼的基因破坏以及斑马鱼的大规模基因组缺失非常有效。在检测的100多个基因组位点中，靶向基因破坏的一次性成功率约为90%，突变频率往往达到50%以上。在这里，我们描述了TALEN组装的验证方案，在热带X线虫和斑马鱼中产生基因敲除动物的方法，以及在斑马鱼中设计大规模基因组缺失的方案。(C)2014 Elsevier Inc.保留所有权利。
Transcription activator like effector nucleases (TALENs) is a promising approach to disrupt intended genomic loci. The assembly of highly effective TALENs is critical for successful genome editing. Recently we reported a convenient and robust platform to construct customized TALENs. The TALENs generated by this platform have been proven to be highly effective for gene disruption in Xenopus tropicalis and zebrafish as well as large genomic deletions in zebrafish. The one-time success rate of targeted gene disruption is about 90% for more than 100 genomic loci tested, with the mutation frequencies often reaching above 50%. Here we describe the validated protocol for TALEN assembly, methods for generating gene knockout animals in X. tropicalis and zebrafish, as well as the protocol for engineering large genomic deletions in zebrafish. (C) 2014 Elsevier Inc. All rights reserved.