Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System.
Homology Directed Knockin of Point Mutations in the Zebrafish tardbp and fus Genes in ALS Using the CRISPR/Cas9 System.
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DOI：
10.1371/journal.pone.0150188
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Drapeau P
中科院分区：
文献类型：
--
作者：
Armstrong GA;Liao M;You Z;Lissouba A;Chen BE;Drapeau P
脊椎动物基因组中单核苷酸的定点编辑方法在生物学和医学研究中具有相当大的兴趣。簇状规则间隔短回文重复序列(CRISPR)/CRISPR相关蛋白9 II型(Cas9)系统已成为一种简单而廉价的工具，用于编辑各种动物模型中感兴趣的基因组位点。在斑马鱼中，易出错的非同源末端连接(NHEJ)已被用作干扰基因功能的一种简单方法。我们试图开发一种方法，轻松地在斑马鱼基因组中创建特定位置的SNPs。在这里，我们报告了使用CRISPR/Cas9介导的同源定向修复的简单方法，使用单链寡核苷酸供体模板(SsODN)进行定点单核苷酸编辑，首次在两个疾病相关基因tardBP和FUS中进行。
The methodology for site-directed editing of single nucleotides in the vertebrate genome is of considerable interest for research in biology and medicine. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 type II (Cas9) system has emerged as a simple and inexpensive tool for editing genomic loci of interest in a variety of animal models. In zebrafish, error-prone non-homologous end joining (NHEJ) has been used as a simple method to disrupt gene function. We sought to develop a method to easily create site-specific SNPs in the zebrafish genome. Here, we report simple methodologies for using CRISPR/Cas9-mediated homology directed repair using single-stranded oligodeoxynucleotide donor templates (ssODN) for site-directed single nucleotide editing, for the first time in two disease-related genes, tardbp and fus.