CRISPR基因编辑技术已成为生命科学研究尤其是人类疾病基因治疗和改造动物进行异种器官移植的利器。然而，文献和我们的前期研究都提示，CRISPR技术会引发动物基因组不稳定性，但这种稳定性改变的特征和规律、遗传稳定性以及不同物种之间的差异性尚不清楚。本研究将通过胚胎分割获得同源胚胎，经CRISPR/Cas9基因编辑制备基因敲除小鼠，对阳性敲除小鼠、空白对照及其F1至F3动物进行深度测序，分析基因组稳定性变化，同时研究靶基因周围基因表达水平及微卫星不稳定性；并用CRISPR基因编辑果蝇、斑马鱼、猪和猴及其同窝阴性对照验证基因组不稳定性变化规律，从横向(不同进化水平)和纵向(不同代数)两个维度探究CRISPR基因编辑技术对基因组稳定性的影响。本研究将为寻求和开发规避遗传风险的CRISPR基因编辑技术提供线索，为其更安全有效地应用于人类疾病治疗和异种器官移植等工作奠定基础。

CRISPR gene editing technology has become a powerful tool for life science researches, especially in gene therapy of human diseases and genetically modified animal xenotransplantation. However, the references and our previous studies indicated that CRISPR technology would induce genomic instabilities in animals. Unfortunately, we still don’t know the characteristics and regulations in such genomic changes, the genetic stability between generations, and the genomic instability among diverse species. In the present study, we will firstly split embryo into homologous embryos. Then knockout mouse would be established by CRISPR/Cas9 system. After the positive knockout mouse generate the offspring, we will collect samples from the positive knockout mouse and its blank control，F1 to F3 to perform genomic sequencing for exploring the genomic instabilities; meanwhile the microsatellite instability and the expression level of the genes closed to the targeted gene will be detected as well. Then the obtained characteristics and regulations of genomic instability will be confirmed by the CRISPR/Cas9 edited drosophila, zebrafishes, pigs and monkeys, and their negative control animals from same litters using genomic sequence. This study will investigate the impacts of the CRISPR gene editing technology on the genomic stability by two dimensions with horizontal (different evolution levels) and vertical (different generations). Our current project could provide clues for the investigation and development of scientific methods for CRISPR technology to avoid genetic risks. We also afford the foundations for the more efficient and safer CRISPR technology by the performance of our project.