Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (37): 6931-6936.doi: 10.3969/j.issn.2095-4344.2012.37.019

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Restraint effect of small interfering RNA targeting UL5 gene on herpes simplex virus type Ⅱ

Pan Xiao-yu, Lü Yan-cheng, Wang Zhi-yong, Fan Jun, Zhou Dan-dan, Yuan Jun-jie   

  1. Department of Biochemistry, Zhuhai Campus, Zunyi Medical College, Zhuhai 519040, Guangdong Province, China
  • Received:2012-01-11 Revised:2012-02-25 Online:2012-09-09 Published:2012-09-09
  • Contact: Lü Yan-cheng, Doctor, Associate professor, Department of Biochemistry, Zhuhai Campus, Zunyi Medical College, Zhuhai 519040, Guangdong Province, China yjskyb@163.com
  • About author:Pan Xiao-yu, Department of Biochemistry, Zhuhai Campus, Zunyi Medical College, Zhuhai 519040, Guangdong Province, China 64262624@qq.com

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Abstract:

BACKGROUND: Helicase-primase complex is the necessary gene for the replication of herpes simplex virus type Ⅱ (HSV-2), and UL5 gene is one of the composing units of HSV-2 helicase-primase complex.
OBJECTIVE: To analyze the interference effect of specific small interfering RNA (siRNA) on HSV-2 UL5 genes using RNA interference technology.
METHODS: To target at HSV-2 UL5 gene, we designed and synthesized five pairs of specific siRNA. By means of liposome LipofeCtamine 2000, the specific siRNA would be transfected into the HEK293 cells. After 48 hours, fluorescence quantitative reverse transcription-PCR test was performed to determine the UL5 gene transcription as well as virus titer detection in order to observe the interference effect of siRNA.
RESULTS AND CONCLUSION: siRNA would be successfully transfected into the cells. The fluorescence quantitative reverse transcription-PCR test showed that siRNA 722, siRNA 2 394, siRNA 2 513 and siRNA 2 627 could variously reduce the expression of the target mRNA. The virus titer detection showed siRNA 722, siRNA 2 394, siRNA 2 513 and siRNA 2 627 could in various degrees lower the titer of virus infection in the supernatant, negative control group and siRNA 374 had no effect on the titer of virus infection. The siRNA which is effective to UL5 gene could specifically reduce the replication of HSV-2.

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