Reagent for CRISPR-Cas9 site-directed mutagenesis editing of DNMT1 gene and application of reagent

The invention discloses a reagent for CRISPR-Cas9 site-directed mutagenesis editing of a DNMT1 gene and application of the reagent. The reagent comprises specific single-stranded guide RNA and an oligodeoxynucleotide template, a target sequence is targeted through a specific single-stranded guide RNA sequence, the oligodeoxynucleotide template is used as a gene repair template, thus, mutation of asingle site is introduced, and single base editing is conducted on the DNMT1 gene to prepare and generate a cell strain carrying DNMT1 site-directed mutagenesis; and the mutated cell strain can be further utilized to study the influence of the DNMT1 mutation on the expression level of a gamma-globin gene (HBG) and the action mechanism of the DNMT1 mutation, and the great application prospects areachieved. 本发明公开了一种用于CRISPR-Cas9定点突变编辑DNMT1基因的试剂及其应用。所述试剂包含特定的单链引导RNA和寡聚脱氧核苷酸模板，通过特定的单链引导RNA序列靶向目标序列，以寡聚脱氧核苷酸模板作为基因修复模板，从而引入单个位点的突变，对DNMT1基因进行单碱基编辑，以制备生成携带该DNMT1定点突变的细胞株；可进一步利用该突变的细胞株来研究该DNMT1突变对γ-珠蛋白基因(HBG)表达水平的影响极其作用机制，具有较大的