中文摘要：

为了揭示IHH（Indian hedgehog）基因在鸡匍匐性状形成过程中的作用机制，利用在线sgRNA平台设计4条IHHsgRNA序列，构建了sgRNA—PX459表达载体，分别命名为sgRNAl、sgRNA2、sgRNA3和sgRNA4。采用脂质体转染至鸡DF-1成纤维细胞后，通过嘌呤霉素筛选4d后收集阳性细胞，采用试剂盒法提取基因组DNA，利用T7E1内切酶和TA克隆测序方法检测4个表达载体的打靶效率。结果表明：4务sgRNA表达载体构建成功，并且都能高效地在DF-1细胞中表达，其中sgRNA1表达载体能有效地打靶IHH基因，通过测序分析发现打靶效率为75．0％，测序结果表明这些突变都以碱基缺失为主。研究结果为揭示IHH基因在鸡匍匐性状中作用机制奠定了坚实的基础。

英文摘要：

In order to properly investigate the mechanism of Indian hedgehog gene （I//H） in regulating creeper phenotype in chicken, four sgRNAs of chicken IHH were designed and corresponding sgRNA-PX459 vectors （sgRNA1, sgRNA2, sgRNA3, and sgRNA4）were also constructed. The four recombinant vectors were then transfected to DF-1 cell by liposomes mediated methods. At 4 days after Puromycin selection, positive cells were collected and genomic DNA was isolated from harvested DF-1 cells by TIANamp Genomic DNA kit （DP304）. A T7E1 digestion method and sequencing of TA cloning were further used to detect the targeting rate of each sgRNA vector. The results showed that four positive recombinant vectors of chicken IHH were gained, and one of them was very efficient in targeting IHH gene. Subsequent sequence analysis showed that the mutation efficiency was approximate 75.0%, and all mutations were mainly expressed in base deletion. The results laid the foundation to reveal the role of IHH gene for creeper phenotype in chicken in the future study.