利用改良的 CRISPR-Cas9 系统在斑马鱼中进行可编程碱基编辑。
Programmable base editing in zebrafish using a modified CRISPR-Cas9 system.
机构信息
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen 518055, China.
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen 518055, China; Department of Molecular, Cell and Development Biology, University of California, Los Angeles, Los Angeles, CA, USA.
出版信息
Methods. 2018 Nov 1;150:19-23. doi: 10.1016/j.ymeth.2018.07.010. Epub 2018 Aug 2.
The use of CRISPR/Cas9 to knockout genes in zebrafish has been well established. However, to better model many human diseases that are caused by point mutations, a robust methodology for generating desirable DNA base changes is still needed. Recently, Cas9-linked cytidine deaminases (base editors) evolved as a strategy to introduce single base mutations in model organisms. They have been used to convert cytidine to thymine at specific genomic loci. Here we describe a protocol for using the base editing system in zebrafish and its application to reproduce a single base mutation observed in human Ablepharon-Macrostomia Syndrome.
CRISPR/Cas9 在斑马鱼中基因敲除的应用已经得到了很好的确立。然而，为了更好地模拟许多由点突变引起的人类疾病，仍然需要一种强大的方法来产生所需的 DNA 碱基变化。最近，Cas9 连接的胞嘧啶脱氨酶（碱基编辑器）作为一种策略在模式生物中引入单碱基突变而得到发展。它们已被用于将胞嘧啶转化为特定基因组位点的胸腺嘧啶。本文描述了在斑马鱼中使用碱基编辑系统的方案及其在复制人类无眼畸形-巨口畸形综合征中观察到的单个碱基突变的应用。
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