Yang Jian, Zhao Chen-Ze, Chen Bin, Chen Fei, Han Jie, Hu Shen-Jiang
Institute of Cardiology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.
Biomed Res Int. 2015;2015:914026. doi: 10.1155/2015/914026. Epub 2015 Jan 22.
Farnesyl pyrophosphate synthase (FPPS) plays a vital role in the mevalonate pathway and has been shown to be involved in hypertrophy and cardiovascular diseases. Lentivirus-mediated RNA interference (RNAi) to knock down a gene of interest has become a promising new tool for the establishment of transgenic animals. The interfering fragment, named pLVT202, was chosen from cardiomyocytes tested in vitro and was microinjected into the perivitelline space of zygotes from C57BL/6J mice via a lentivirus vehicle; 20 were identified as carrying copies of the transgene using the polymerase chain reaction (PCR). Real-time PCR and western blotting analysis showed that FPPS was downregulated in multiple tissues in the transgenic mice. The transgenic mouse model provides a novel means of studying the gene function of FPPS.
法尼基焦磷酸合酶(FPPS)在甲羟戊酸途径中起着至关重要的作用,并且已被证明与肥大和心血管疾病有关。慢病毒介导的RNA干扰(RNAi)敲低感兴趣的基因已成为建立转基因动物的一种有前景的新工具。从体外测试的心肌细胞中选择干扰片段pLVT202,并通过慢病毒载体将其显微注射到C57BL/6J小鼠受精卵的卵周隙中;使用聚合酶链反应(PCR)鉴定出20只携带转基因拷贝。实时PCR和蛋白质印迹分析表明,转基因小鼠的多个组织中FPPS表达下调。该转基因小鼠模型为研究FPPS的基因功能提供了一种新方法。