中国组织工程研究Issue(29)：4605-4610,6.DOI:10.3969/j.issn.2095-4344.2014.29.003
HLA-A2小干扰RNA的设计及沉默效果检测
Designation and silencing function of the small interfering RNA of HLA-A2
摘要
Abstract
BACKGROUND:Human leukocyte antigen (HLA), the major histocompatibility complex of human, plays an important function in the transplant rejection. Decreasing the expression of HLA wil prolong the survival time of transplants. <br> OBJECTIVE:To design smal interfering RNA (siRNA) of HLA-A2 and to detect the effect of siRNA-HLA-A2 on the expression of HLA-A2. <br> METHODS:Four kinds of siRNA-HLA-A2 domains were designed, and recombinant lentivirus expression vector were formed. The 293T cells, highly expressing HLA-A2, were infected in vitro. Then the knockout efficacy of four domains was detected to select the highly efficient siRNA-HLA-A2 target sequences. The human embryo lung fibroblasts were cultured in vitro and infected with the lentivirus carrying the target sequence. The infecting efficiency of LV-siRNA-HLA-A2 was observed under the fluorescence microscope and the silence function of this siRNA in human embryo lung fibroblasts was detected by western blot analysis. <br> RESULTS AND CONCLUSION:According to the mRNA sequence of HLA-A2 in Genbank, three siRNAs were designed and synthesized. In vitro, the over expression of HLA-A2 in 293K cells was successful y silenced. The HLA-A2 expression in human embryo lung fibroblasts was also efficiently silenced after the human embryo lung fibroblasts were infected by the highly efficient siRNA of HLA-A2. The efficacy was up to 80%.关键词
组织构建/组织工程/人类白细胞抗原A2/RNA干扰/基因沉默/人胚肺成纤维细胞/辽宁省自然科学基金Key words
HLA antigens/RNA interference/gene silencing/fibroblasts分类
医药卫生引用本文复制引用
刘建生,赵刚,裴丹,郑德宇..HLA-A2小干扰RNA的设计及沉默效果检测[J].中国组织工程研究,2014,(29):4605-4610,6.基金项目
辽宁省自然科学基金资助项目(2013022066，2013022048)；辽宁省教育厅项目(L2013335)；辽宁医学院青年基金项目(Y2011Z022)@@@@the Natural Science Foundation of Liaoning Province, No.2013022066,2013022048 （2013022066，2013022048）
a grant by Liaoning Provincial Education Bureau, No. L2013335 （）
the Youth Fund of Liaoning Medical University, No. Y2011Z022 （）
