Liu Ji-feng, Guan Cui-ping, Tang Xu, Xu Ai-e
Department of Dermatology, Third Hospital of Hangzhou, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2010 Jun;24(3):199-201.
To explore the inhibition effect of RNA interference on the ICP4 expression and DNA replication of herpes simplex virus type 2 (HSV2).
Four pairs of siRNA targeted to HSV2 ICP4 gene and negative control siRNA were synthetized by chemical method, named as siRNA-1, siRNA-2, siRNA-3, siRNA-4 and siRNA-N respecticely. HSV2 HG52 was used to attack Vero cell after transfection overnight. Vero cell and supernatant were collected at 1d, 2d, 3d, 4d and 5d after virus attacking. Flurogenic quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR)was used to detect the expression of HSV2 ICP4 mRNA, flurogenic quantitative polymerase chain reaction(FG-PCR) was used to detect the expression of HSV2 DNA and Western-Blot was used to detect the expression of HSV2 ICP4 protein.
All the four pairs of siRNA could significantly inhibit the expression of HSV2 ICP4 mRNA and protein, especially siRNA-2. The above siRNAs could significantly decrease HSV2 DNA copy number,too.
siRNAs targeted to HSV2 ICP4 gene could significantly inhibit expression of HSV2 ICP4 mRNA and protein, and decrease HSV2 DNA copy number, suggesting that siRNA can inhibit HSV2 DNA replication through silencing ICP4 gene.
探讨RNA干扰对2型单纯疱疹病毒（HSV2）ICP4表达及DNA复制的抑制作用。
采用化学合成法合成4对针对HSV2 ICP4基因的小干扰RNA（siRNA）及阴性对照siRNA，分别命名为siRNA-1、siRNA-2、siRNA-3、siRNA-4和siRNA-N。转染过夜后，用HSV2 HG52攻击Vero细胞。病毒攻击后1天、2天、3天、4天和5天收集Vero细胞及上清。采用荧光定量逆转录聚合酶链反应（FQ-RT-PCR）检测HSV2 ICP4 mRNA的表达，荧光定量聚合酶链反应（FG-PCR）检测HSV2 DNA的表达，蛋白质免疫印迹法（Western-Blot）检测HSV2 ICP4蛋白的表达。
4对siRNA均能显著抑制HSV2 ICP4 mRNA和蛋白的表达，其中siRNA-2作用尤为明显。上述siRNAs也能显著降低HSV2 DNA拷贝数。
靶向HSV2 ICP4基因的siRNAs能显著抑制HSV2 ICP4 mRNA和蛋白的表达，并降低HSV2 DNA拷贝数，提示siRNA可通过沉默ICP4基因抑制HSV2 DNA复制。
