摘要
为了研究RNA干扰(RNAi)对Ⅰ型登革病毒(DENV-1)在白纹伊蚊C6/36细胞内复制的影响,本研究设计并合成针对I型登革病毒Pr M基因的小干扰RNA,以脂质体法转染入C6/36细胞后,用DENV-1感染已转染的细胞,观察细胞病变效应,MTT法检测细胞存活率,荧光定量RT-PCR检测登革病毒RNA含量。结果表明:转染siRNA的C6/36细胞在受登革病毒攻击7天后仍无明显细胞病变效应,细胞存活率比对照组提高2.26倍,细胞内登革病毒RNA拷贝数比对照组降低约97.54%。说明利用RNA干扰技术能有效抑制登革病毒核酸在C6/36细胞内复制,并对细胞具有一定保护作用,为登革热的防治提供了新的思路。
To investigate the inhibitory effect of RNA interference（RNAi） on dengue virus Ⅰ（DENV-1） replication.Small interfering RNA（siRNA） against the PreM gene of dengue virus was synthesized and transfected into C6/36 cells with liposome,which was then attacked by DENV-1 virus.The antiviral effect of siRNA was evaluated by cytopathic effect（CPE）,the cell survival rate measured by MTT,and virus RNA quantified by real-time RT-PCR.The results showed that after 7 days post infection of dengue virus,the transfected C6/36 cells showed less CPE.The cell survival rate of the transfected C6/36 cells increased by 2.26 fold,and the amount of virus RNA in the transfected cells was reduced by about 97.54% as well.These findings indicated that the siRNA could effectively inhibit dengue virus RNA replication,and protect C6/36 cells from viral attack,indicating its potential role in prevention and treatment of dengue fever.
出处
《病毒学报》
CAS
CSCD
北大核心
2010年第5期373-378,共6页
Chinese Journal of Virology
基金
广东省科技计划项目(2008B030303041)
广州市科技计划项目(2006J1-C0141
2008J1-C191
2008Z1-E231)
广州市医学科学技术研究重点项目(2006-ZDi-10
2008-ZDi-12)
