中国组织工程研究2012，Vol.16Issue(7)：1267-1270,4.DOI:10.3969/j.issn.1673-8225.2012.07.030
细胞因子信号转导抑制因子1RNA干扰慢病毒载体的构建与鉴定
Construction and identification of a lentiviral vector for RNA interference of suppressors of cytokine signaling 1
摘要
Abstract
BACKGROUND: The previous studies mainly used liposome transfection or other vectors for research of suppressors of cytokinesignaling 1 (SOCS1). However, those transfections have the disadvantages of low efficiency and less security.OBJECTIVE: To construct a lentiviral vector for RNA interference of SOCS1 gene and identify it.METHODS: Three pairs of complementary short hairpin RNA (shRNA) oligonucleotides targeting the SOCS1 gene weredesigned, and then were inserted into pPll3.7 vectors by gene recombination technology. The recombinant plasmids with correctsequence were cotransfected with packaging plasmids into 293T cells. The cell culture supernatant was obtained after 48 hoursin order to transfect A549 cells. The expression of SOCS1 in target cells was detected by western-blot.RESULTS AND CONCLUSION: The shRNA had been confirmed to insert into pPll3.7 vectors by sequencing of the recombinantplasmids. The western-blot detection showed that the expression of SOCS1 in A549 cells was inhibited after the cells weresuccessfully transfected by the lentiviral vector supernatant. A lentiviral vector for RNA interference of SOCS1 gene wassuccessfully constructed in this study.关键词
细胞因子信号转导抑制因子1/慢病毒载体/RNA干扰/A549/细胞/组织构建分类
医药卫生引用本文复制引用
周敏,钱嫣蓉,张秋蕊,程挺,万欢英..细胞因子信号转导抑制因子1RNA干扰慢病毒载体的构建与鉴定[J].中国组织工程研究,2012,16(7):1267-1270,4.基金项目
上海市科委医药处重点项目( 编号09411951400),项目名称:SOCS1 在特发性肺纤维化中临床意义的研究. （ 编号09411951400）
