Lentviral-mediated RNAi to inhibit target gene expression of the porcine integrin alpha v subunit, the FMDV receptor, and against FMDV infection in PK-15 cells

文献类型： 外文期刊

第一作者： Luo, Jihuai

作者： Luo, Jihuai;Du, Junzheng;Gao, Shandian;Zhang, Guofeng;Sun, Jingjing;Cong, Guozheng;Shao, Junjun;Lin, Tong;Chang, Huiyun

作者机构：

期刊名称：VIROLOGY JOURNAL （ 影响因子：4.099； 五年影响因子：3.719 ）

ISSN： 1743-422X

年卷期： 2011 年 8 卷

页码：

收录情况： SCI

摘要： Background: shRNA targeting the integrin alpha v subunit, which is the foot-and-mouth disease virus (FMDV) receptor, plays a key role in virus attachment to susceptible cells. We constructed a RNAi lentiviral vector, i alpha v pLenti6/BLOCK -iT (TM), which expressed siRNA targeting the FMDV receptor, the porcine integrin av subunit, on PK-15 cells. We also produced a lentiviral stock, established an i alpha v-PK-15 cell line, evaluated the gene silencing efficiency of mRNA using real-time qRT-PCR, integrand alpha v expression by indirect immunofluorescence assay (IIF) and cell enzyme linked immunosorbent assays (cell ELISA), and investigated the in vivo inhibitory effect of shRNA on FMDV replication in PK-15 cells. Results: Our results indicated successful establishment of the i alpha v U6 RNAi entry vector and the iav pLenti6/BLOCK-iT expression vector. The functional titer of obtained virus was 1.0 x 10(6) TU/mL. To compare with the control and mock group, the i alpha v-PK-15 group alpha v mRNA expression rate in group was reduced by 89.5%, whilst IIF and cell ELISA clearly indicated suppression in the experimental group. Thus, i alpha v-PK-15 cells could reduce virus growth by more than three-fold and there was a > 99% reduction in virus titer when cells were challenged with 102 TCID50 of FMDV. Conclusions: I alpha v-PK-15 cells were demonstrated as a cell model for anti-FMDV potency testing, and this study suggests that shRNA could be a viable therapeutic approach for controlling the severity of FMD infection and spread.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序