丙型肝炎病毒特异性核酶对HepG2.9706细胞HCV5′NCR基因表达的抑制作用

 为探讨锤头型核酶抗丙型肝炎病毒的作用 ,根据HCV 5′NCR及翻译起始区的二级结构 ,设计及合成了 1个锤头型核酶RzA .将其插入pCI neo表达载体的多克隆位点 ,构建了表达RzA的质粒pHCV RzA .采用转基因细胞模型HepG2 970 6细胞 ,评价了pHCV RzA质粒对HCV 5′NCR调控荧光素酶表达的抑制活性 .结果表明 ,在HepG2 970 6细胞中 ,pHCV RzA以序列特异性、剂量依赖性方式抑制荧光素酶基因的表达 ,抑制率达 80 % ,提示核酶有可能作为HCV感染基因治疗的一种有效途径 .

The Inhibitory Effects of Gene Expression Controlled by HCV 5'NCR in HepG2.9706 by Hammerhead Ribozyme

Ribozymes are RNA molecules that catalyze cleavage of a target RNA molecule based on sequence specific recognition To determine the effects of hammerhead ribozyme against hepatitis C virus(HCV)on viral protein translation,a hammerhead ribozyme RzA directed to the translation initiation region of HCV RNA was designed and synthesized according to the secondary structure of HCV 5′NCR and adjacent C region RzA gene was inserted into multiple cloning sites of pCI neo vector and constructed the RzA producing plasmid(pHCV RzA) HepG2 9706 cells were transfected using pHCV RzA via Lipofectin for three consecutive days The results showed that pHCV RzA inhibits luciferase expression,controlled by HCV 5′NCR in HepG2 9706 cells,by 48%-80% The results demonstrate the potential of ribozyme therapy in the treatment of HCV infection