转座子是染色体上一种可以改变位置的基因，可以实现不同位置之间的转换，也可以从外源DNA转移到宿主基因组中^[1]，而转座子编码出来的转座酶则可以通过特异性识别转座子两端的序列，使其从原位点脱离出来，然后整合到新的靶位点^[2]。ASS1基因的一些碱基突变可能会导致一些疾病的发生。

本实验主要是想要探究转座子与CRISPR/Cas9基因编辑技术相结合对ASS1相关遗传病的应用。实验中通过CRISPR/Cas9技术构建打靶质粒，通过细胞转染，使293ft细胞中的ASS1基因发生突变，提取基因组后，进行T7酶切检验打靶效率；通过PCR技术从细胞中获得PB基因，进行同源重组，获得PB质粒，用于后续实验的工具构建，前期准备工作还算成功。

Transposon is a kind of gene can change the position on the chromosome, can realize the conversion between different positions, can also be transferred from foreign DNA to the host genome^[1], and transposon encoded the transposon can specifically identify the ends of the transposon, make it from the original site, and then integrated into a new target site^[2]. Some base mutations in the ASS1 gene may lead to the generation of some diseases.

This experiment was mainly designed to explore the application of transposons combined with CRISPR/Cas 9 gene editing technology to ASS1-related genetic diseases. In the experiment, the target plasmid was constructed by CRISPR/Cas 9 technology, and the ASS1 gene in 293ft cells was mutated by cell transfection. After the genome was extracted, T7 digestion tested the target efficiency. PB gene was obtained from cells by PCR technology, homologous recombination, and PB plasmid was obtained for tool construction of subsequent experiments, and the preparatory work was successful.