利用RNA干扰技术抑制单胺氧化酶B基因的表达

Inhibition of Monoamine Oxidase B Expressions in Hela Cell by siRNA

目的确定并筛选用于特异静默单胺氧化酶B(Monoamine Oxidase B,MAOB)基因的干扰小RNA(siRNA)片段,建立RNA干扰技术平台。方法构建含单胺氧化酶B基因siRNA片段的pSilencer 1.0-U6 siRNA表达质粒,转染Hela细胞,用RT-PCR和定量PCR方法检测单氨氧化酶B mRNA的水平。结果所设计的两段siRNA片段均有干扰MAOB mRNA水平的作用,片段2的作用优于片段1,两个片段之间有一定的协同作用。结论选择的siRNA片段可以抑制Hela细胞单胺氧化酶B的表达。

Objective To select specific siRNA sequences that can silence the MAOB gene. Methods Inserting MAOB specific siRNA sequence into pSilencer 1.0-U6 siRNA expressing vector and then transfecting the vector into Hela cells. The MAOB mRNA transcription were examined by RT-PCR and real time PCR. Results Two sequences were tested and both showed RNAi effects. But the inhibition efficiency of Sequence 2 was better. A higher silencing rate was observed when two vectors were co-transfected. Conclusion The selected sequences are useful for the silencing of MAOB gene.