瞬时转染siRNA抑制大鼠神经干细胞内MMS2基因的表达  

Inhibition of transient transfection siRNA on MMS2 expression in rat neural stem cells

作　　者：冯海娇[1] 沈岳飞[1] 罗小丹[2] 罗彦妮[3] 

机构地区：[1]广西医科大学第一附属医院神经内科,南宁市530021 [2]广西钦州市第二人民医院神经内科,钦州市535000 [3]广西民族医院神经内科,南宁市530001

出　　处：《微创医学》2011年第1期4-9,共6页Journal of Minimally Invasive Medicine

基　　金：广西科学基金(编号:桂科自0832140)

摘　　要：目的建立有效的脂质体瞬时转染法,转染小分子干扰RNA(small interfering RNA,siRNA),沉默大鼠神经干细胞(neural stem cells,NSCs)内甲基磺酸敏感蛋白2(Methyl Methanesulfonate Sensitive 2,MMS2)的表达。方法设计并化学合成针对MMS2基因的3对siRNA分子序列,采用阳离子脂质体法瞬时转染NSCs,运用实时荧光定量PCR检测NSCs内MMS2基因mRNA的表达。结果 MMS2-siRNA对大鼠NSCs内MMS2基因的沉默效果在转染后36 h达到最大抑制作用;3对特异性MMS2-siRNA序列均有效地抑制了MMS2基因的表达,沉默效率分别为siRNA_A(58±5)%、siRNA_B(64±6)%、siRNA_C(84±3)%,与空白对照组相比均存在显著性差异(P<0.05);siRNA_C沉默效率最高。结论靶向MMS2基因的siRNA分子片段可以有效地抑制NSCs内MMS2基因的表达,为后续相关研究提供实验方法。Objective To explore the effective method of transfecting small interfering RNA（siRNA） to mute the expression of methyl methanesulfonate sensitive 2（MMS2） in rat neural stem cells（NSCs） by liposomes instant transfection.Methods Three siRNAs were designed and synthesized according to the mRNA sequence of rat MMS2 gene,and transfected into NSCs in culture by Cationic liposomes.To assess the efficiency of siRNA knockdown,the gene expression of MMS2 was detected by real-time PCR.Results The maximum inhibitory effects of MMS2-siRNA transfection on MMS2 expression was at 36h.The MMS2 gene was knocked down by siRNA-A,siRNA-B,and siRNA-C,with silence rate of（58±5）%,（64±6）%,and（84±3）%,respectively.siRNA_C was the most effective siRNA.Conclusion The MMS2 gene in rat NSCs can be knocked down effectively by RNAi,serving as a method for the consequential experiment.

关 键 词：RNA干扰 神经干细胞 甲基磺酸敏感蛋白2 阳离子脂质体 

分 类 号：R742.5[医药卫生—神经病学与精神病学]