β位淀粉样前体蛋白裂解酶2基因敲除斑马鱼动物模型的构建及其初步应用

Construction and preliminary application of BACE2 gene knockout in zebrafish using CRISPR / Cas9

目的构建β位淀粉样前体蛋白裂解酶2(β-site APP-cleaving enzyme 2,BACE2)基因敲除斑马鱼动物模型并及其初步探索其应用。方法运用CRISPR/Cas9技术,在斑马鱼中设计针对靶点的gRNA,使之诱导靶点突变,经测序鉴定得到突变体。结果BACE2纯合突变体在早期部分无法存活,RT-PCR结果显示BACE2基因在斑马鱼的早期发育中持续表达。结论 BACE2基因敲除的斑马鱼动物模型将为进一步研究BACE2基因在斑马鱼胰腺发育中的作用和针对糖尿病的药物靶点筛选提供有效工具。

Objective To construct β-site APP-cleaving enzyme 2 knockout zebrafish animal model and preliminarily explore its application. Methods By using the CRISPR / Cas9 technology,the gRNA against target in zebrafish was designed to induce mutations;afterwards the mutant was obtained by DNA sequencing. Results BACE2 homozygous mutants were unable to survive in the early part.Results of RT-PCR showed continued BACE2 gene expression in zebrafish early development. Conclusion BACE2 knockout zebrafish animal model may provide an effective tool for further study of the mechanism of pancreas development and drug screening.