Abstract: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, doublestranded DNA of the Asb11 gene targeting gRNA was amplified by PCR. Thirdly, the Asb11 gene gRNA and Hcas9 mRNA were coinjected into zebrafish embryos at 1cell stage. The results of the effectivenesstest 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 knockedout were established successfully. The work laid a foundation for exploring the roles of Asb11 in zebrafish heart development.
引用本文:
尹丽阳,罗世锋,陈〓宇,漆轲婧,彭〓云,万永奇,吴秀山,李永青. 利用CRISPR/Cas9 技术建立斑马鱼Asb11基因敲除品系[J]. 激光生物学报, 2018, 27(3): 252-259.
YIN Liyang, LUO Shifeng, CHEN Yu, QI Kejing, PENG Yun, WAN Yongqi, WU Xiushan, LI Yongqing. Establishing the Asb11 Knockout Lines of Zebrafish with CRISPR/Cas9 Targeting Technology. journal1, 2018, 27(3): 252-259.