CRISPR/Cas9 and TALEN-mediated knock-in approaches in zebrafish
CRISPR/Cas9 and TALEN-mediated knock-in approaches in zebrafish
复制标题
DOI：
10.1016/j.ymeth.2014.03.027
复制
发表时间：
2014-09-01
期刊：
影响因子：
4.8
通讯作者：
Del Bene, Filippo
中科院分区：
文献类型：
--
作者：
Auer, Thomas O.;Del Bene, Filippo
利用序列特异性转录激活子样效应核酸酶 (TALEN) 和成簇规则间隔短回文重复序列 (CRISPR)/CRISPR 相关 (Cas) 9 系统（RNA 引导核酸酶，RGN）定向引入突变，彻底改变了许多模式生物中的反向遗传方法。在斑马鱼中，这两个系统都成功应用于通过靶向开放阅读框或整个染色体区域的删除和倒位来生成功能丧失的等位基因。除了产生这些功能丧失的等位基因之外，通过利用单链 DNA 寡核苷酸作为基于同源性修复的模板插入短序列的基因组工程也成为可能，从而能够有效插入蛋白质编码基因的 loxP 位点或标签。最近基于同源重组和非同源末端连接的研究也扩大了基因组编辑的范围。这些方法允许有针对性地插入开放阅读框甚至整个供体载体。在这篇综述中，我们总结了 TALEN 和 RNA 引导的核酸酶在斑马鱼遗传学领域的应用，特别关注敲入方法。 (C) 2014 Elsevier Inc. 保留所有权利。
The targeted introduction of mutations utilizing sequence specific transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9 system (RNA-guided nucleases, RGNs) has revolutionized reverse genetic approaches in numerous model organisms. In zebrafish, both systems were successfully applied to generate loss-of-function alleles by targeting open reading frames or deletion and inversion of whole chromosomal regions. In addition to the production of these loss-of-function alleles, genomic engineering by insertion of short sequences utilizing single stranded DNA oligonucleotides as templates for homology based repair was made possible, enabling effective insertion of loxP sites or tags for protein coding genes. Recent studies based on homologous recombination and non-homologous end joining have also broadened the repertoire for genome editing. These approaches allow the targeted insertion of open reading frames or even whole donor vectors. In this review we summarize the use of TALENs and RNA-guided nucleases in the field of zebrafish genetics with a special focus on knock-in approaches. (C) 2014 Elsevier Inc. All rights reserved.