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MeCP2(Methyl CpG binding protein 2)基因突变可导致Rett综合征(Rett syndrome,RTT)。目前已报道的MeCP2敲除小鼠表型与RTT病人症状存在显著差异。为探索MeCP2在脑发育中的作用及其导致RTT的机制,本研究利用CRISPR/Cas9技术构建了MeCP2基因敲除大鼠模型。通过构建靶向敲除MeCP2基因的载体,体外将Cas9 mRNA和sgRNA显微注射到SD大鼠受精卵中,在MeCP2基因exon2中造成移码突变,从而获得MeCP2基因敲除大鼠。利用测序和Westernblotting方法鉴定MeCP2敲除大鼠,并对其表型和行为学特征进行分析,发现MeCP2敲除大鼠体重降低,存在焦虑倾向和认知缺陷。本研究成功构建了MeCP2基因敲除大鼠模型,其表型类似人类RTT患者的症状,为后续MeCP2功能研究提供了更好的动物模型。
MeCP2 (Methyl CpG binding protein 2) gene mutation can cause Rett syndrome (Rett syndrome, RTT). There are significant differences between the phenotype of MeCP2 knockout mice and the symptoms of RTT that have been reported so far. In order to explore the role of MeCP2 in brain development and its mechanism leading to RTT, we constructed a rat model of MeCP2 knockout using CRISPR / Cas9 technology. By constructing a vector targeting MeCP2 gene knockout, Cas9 mRNA and sgRNA were microinjected into the zygotes of SD rats in vitro, resulting in frame-shifting mutations in exon2 of MeCP2 gene, thereby obtaining MeCP2 knockout rats. The MeCP2 knockout rats were identified by sequencing and Western blotting. The phenotypic and behavioral characteristics of MeCP2 knockout rats were analyzed. It was found that MeCP2 knockout rats had reduced body weight, anxiety and cognitive deficits. In this study, MeCP2 gene knockout rat model was successfully constructed, and its phenotype resembled the symptoms of human RTT patients, which provided a better animal model for the subsequent study of MeCP2 function.