中文摘要：

斑马鱼是研究人类疾病的重要动物模型,基因组定点编辑技术在利用斑马鱼研究同源基因功能方面提供了简便高效的途径。利用CRISPR/Cas9系统,在斑马鱼TU品系第5号染色体中选择靶位点,将Cas9 mRNA、靶位点识别的sgRNA和带有同源重组臂、利用短链肌球蛋白（Mylz2）启动子启动的绿色荧光蛋白（eGFP）基因显微注射到斑马鱼受精卵中,获得在该位点定点插入外源基因,并在肌肉中特异表达绿色荧光蛋白的转基因斑马鱼。利用基因组DNA的PCR扩增,结合激发光下斑马鱼胚胎荧光表达情况,在存活的87尾幼鱼中发现其中45尾有荧光表达,同源重组率为51.724%。研究表明,利用CRISPR/Cas9系统成功定点插入外源基因,并获得在肌肉中特异性表达绿色荧光的斑马鱼。

英文摘要：

Zebrafish is an important animal model for studying human diseases. Gene editing is an important research tool for the improvement of animal breeds and the establishment of animal models of human diseases and gene editing provides a convenient and efficient way to study the functions of genes in zebrafish. Using the CRISPR/Cas9 system,Cas9 mRNA,a fragment containing homologous recombination arms and green fluorescent protein（ EGFP） gene driven by short chain myosin（ Mylz2） gene promoter and sgRNA,which targets to chromosome 5 of TU strain,were micro-injected into the zebrafish fertilized eggs. Transgenic zebrafish were identified by PCR of the genomic DNA accompanied with the observation of green fluorescent protein under microscopy. The results showed that 45 of 87 survival fish gained homologous recombination with a recombination rate of 51. 724%. The study indicated that a foreign eGFP gene was successfully inserted into zebrafish genome and was specifically expressed in the muscle using CRISPR/Cas9 system.