Abstract: Objective To construct a recombinant lentiviral vector carrying miR-1908 and validate its infection efficiency in human pre-adipocytes. Methods The minigene fragments of miR-1908 was amplified from genomic DNA of human pre-adipocytes by PCR,then cloned into a lentivirus expression vector.After sequencing validated exactly,the expression plasmid and packaging plasmids were co-transfected into HEK-293T cells to produce lentivirus;The supernatant containing lentiviral particles was used to infect human pre-adipocytes,and its infection efficiency and miR-1908 overexpression level were respectively tested by fluorescent observation and real-time quantitative PCR. Results miR-1908 lentiviral vector was constructed successfully,and infection efficiency in human pre-adipocytes reached over 70%,the expression level increased nearly two folds in infected cells. Conclusion It has been successfully constructed that the lentiviral vector containing miR-1908 with high overexpression in infected human pre-adipocytes,which will be useful to the future studying the function of miR-1908 in human pre-adipocytes

Key words: miR-1908, lentivirus, human preadipocytes

摘要： 目的 构建人miR-1908慢病毒表达载体，包被病毒并检测其在人脂肪前体细胞中过表达效果。方法 以人脂肪细胞基因组DNA为模板，PCR高保真扩增包含miR-1908区域上下游各100bp左右的基因组DNA，亚克隆入慢病毒表达载体，与包装质粒共转染HEK-293T细胞，包装病毒，收取病毒上清，纯化后感染人脂肪前体细胞，36 h开始观察荧光标记，72 h收取细胞，抽提RNA，Realtime PCR检测慢病毒感染下miR-1908的相应表达量。结果 成功构建人miR-1908慢病毒表达载体，包装获得的病毒感染人脂肪前体细胞的效率可达到70%以上，miR-1908过表达水平可接近2倍。结论 本研究成功构建了人miR-1908慢病毒过表达载体，包被的慢病毒可以在人脂肪前体细胞中实现过表达效果，为后续功能研究奠定了基础。

关键词: miR-1908, 慢病毒, 人脂肪前体细胞