Asb11基因被报道与斑马鱼Notch信号的激活有关，本研究室过去的研究显示该基因在心肌和骨骼肌中特异性表达。因此推测Asb11基因可能是心脏发育相关候选基因。为了阐明Asb11基因在斑马鱼心脏发育过程中的作用，本文利用CRISPR/Cas9打靶技术构建敲除Asb11基因的斑马鱼品系。首先在线分析筛选出Asb11基因最适合的打靶位点，然后PCR扩增出Asb11基因gRNA的双链cDNA，再将Asb11基因的gRNA和Hcas9的mRNA共同注射到斑马鱼胚胎Ⅰ细胞期胚胎中。进行打靶的有效性检测，发现Asb11基因的一号外显子出现了碱基的缺失，表明CRISPR/Cas9 系统对Asb11基因的敲除是有效的。对其F0代、F1代、F2代进行筛选，成功获得了Asb11基因敲除的斑马鱼品系，为探究Asb11在心脏发育中的作用奠定了基础。

Asb11 gene was reported that it was associated with activation of Notch signaling pathway. The previous results in our lab showed that the gene expressed specifically in cardiac muscle and skeletal muscle. Therefore, it was hypothesized that Asb11 gene might be a candidate gene related to heart development. In this study, CRISPR/Cas9 targeting technology was employed to knockout zebrafish Asb11 gene. Firstly, the most suitable target sites in Asb11 gene were screened out on websites. Secondly, doublestranded DNA of the Asb11 gene targeting gRNA was amplified by PCR. Thirdly, the Asb11 gene gRNA and Hcas9 mRNA were coinjected into zebrafish embryos at 1cell stage. The results of the effectivenesstest showed that bases were deleted in the exon1 of the zebrafish Asb11 gene, suggesting that the CRISPR/Cas9 system was effective for knockout of Asb11 gene. After screening in the F0, F1 and F2 generations, the lines of zebrafish with Asb11 gene knockedout were established successfully. The work laid a foundation for exploring the roles of Asb11 in zebrafish heart development.