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.

中文翻译：

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使用改良的 CRISPR-Cas9 系统在斑马鱼中进行可编程碱基编辑

使用 CRISPR/Cas9 敲除斑马鱼中的基因已经很成熟。然而，为了更好地模拟由点突变引起的许多人类疾病，仍然需要一种强大的方法来产生理想的 DNA 碱基变化。最近，Cas9 连接的胞苷脱氨酶（碱基编辑器）进化为一种在模式生物中引入单碱基突变的策略。它们已被用于在特定基因组位点将胞苷转化为胸腺嘧啶。在这里，我们描述了在斑马鱼中使用碱基编辑系统的协议及其在人类 Ablepharon-Macrostomia 综合征中观察到的单个碱基突变的应用。