Abstract：Abstract: As a member of the WW domain binding protein family, WBP11 is an important splicing factor involved in the pre-mRNA splicing process. Zebrafish, one of the notable model organisms, contains orthological homologous gene wbp11 in its genome. In order to study the mechanism of WBP11 in the early development in vertebrates, a knockout strain of wbp11 in zebrafish was constructed using CRISPR/Cas9 technology. Firstly, gene knockout target sites were designed on the second exon of wbp11 gene, and sgRNA was synthesized by transcription in vitro. After microinjection technology, F0 generation chimeric zebrafish with wbp11-knockout was obtained. Then, the F1 generation zebrafish by crossing chimeric zebrafish with wild-type zebrafish were identified by genotype, and heterozygous zebrafish were screened out. The sequencing results for exact bands of heterozygous zebrafish showed that there were two strains with different sequence deletion. Heterozygous zebrafish of the F1 generation of the same strain were self-crossed. The homozygous zebrafish was screened out via genotyping, meaning the wbp11 gene knockout strain was successfully constructed. The zebrafish with 48 hours post fertilization showed pericardial edema, heart linearization and skeletal curvature obversed by microscopic white light imaging. These defections may be resulted from the abnormal splicing mechanism caused by the deletion of wbp11 gene. This study opens the way to explore the mechanism of WBP11 gene in the early development in vertebrates. Key words: zebrafish; wbp11; CRISPR/Cas9; gene knockout; heterozygote （Acta Laser Biology Sinica, 2023, 32(3): 217-225）
引用本文:
游诗琦，陈 宇，杨雪婷，李韵璇，乔 卿，罗 莹，姚梅英，高罗晴，刘 欣，王跃群，吴秀山，李永青. 斑马鱼wbp11基因敲除品系的建立[J]. 激光生物学报, 2023, 32(3): 218-225. YOU Shiqi, CHEN Yu, YANG Xueting, LI Yunxuan, QIAO Qing, LUO Ying, YAO Meiying, GAO Luoqing, LIU Xin, WANG Yuequn, WU Xiushan, LI Yongqing. Construction of Zebrafishwbp11Gene Knockout Line. journal1, 2023, 32(3): 218-225.
