利用2型重组腺相关病毒抑制子宫腔上皮Plekhs1基因表达对小鼠生育能力的影响  

Effect of uterine epithelial-specific Plekhs1 knockdown mediated by recombinant adeno-associated virus type 2(rAAV2)on mouse fertility

作　　者：马兴红[1] 张其法 姜南 李世杰[1] 王一妹 MA Xinghong;ZHANG Qifa;JIANG Nan;LI Shijie;WANG Yimei(School of Life Sciences,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《东北农业大学学报》2020年第6期70-78,共9页Journal of Northeast Agricultural University

基　　金：国家自然科学基金项目(31571553)。

摘　　要：研究构建稳定表达Plekhs1 shRNA 2型重组腺相关病毒载体(rAAV2-GFP-shRNA-Plekhs1),筛选并制备抑制Plekhs1表达效果最强重组腺相关病毒(rAAV2-PL3)。重组病毒子宫角注射到小鼠子宫,通过Real-time PCR、荧光检测、免疫组化技术检测rAAV2-PL3在小鼠子宫侵染表达规律,分析rAAV2-PL3侵染小鼠子宫抑制Plekhs1表达效果及对生殖影响。结果表明,rAAV2-PL3可有效转染至小鼠子宫内膜上皮细胞中并抑制Plekhs1表达,F1代产仔数量显著降低。研究表明,子宫角注射重组腺相关病毒可高效研究子宫腔上皮基因功能。In this study,a type 2 recombinant adeno-associated virus vector(rAAV2-GFPshRNAPlekhs1)stably expressing Plekhs1 shRNA was constructed,and a recombinant adeno-associated virus(rAAV2-PL3)with the strongest inhibitory effect on Plekhs1 expression was screened and prepared.After the recombinant virus was injected into the mouse uterus,the expression of rAAV2-PL3 in the mouse uterus was detected by real-time PCR,fluorescence detection,and immunohistochemistry.The effect of rAAV2-PL3 on the inhibition of Plekhs1 expression and reproduction were analyzed.The results showed that rAAV2-PL3 could be effectively transfected into mouse endometrial epithelial cells,inhibited the expression of Plekhs1,and resulted in a significant reduction in the number of F1 generation litters.These findings demonstrated that the uterine injection of recombinant adenoassociated virus(rAAV)could efficiently study the gene function of uterine cavity epithelium.

关 键 词：AAV2 基因转导 生殖发育 小鼠子宫 

分 类 号：Q331[生物学—遗传学]