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Lentviral-Mediated RNAi for the Control of Foot and Mouth Disease Virus

8 November 2011, at 12:00am

The interference integrin av subunit expressing system is a feasible strategy for overcoming the issue of high genetic variability of the foot and mouth disease virus (FMDV) and viral escape, report researchers from China.

shRNA targeting the integrin av subunit, which is the FMDV receptor, plays a key role in virus attachment to susceptible cells, according to Jihuai Luo and colleagues at the Chinese Academy of Agricultural Sciences in Lanzhou. In the introduction to their paper published in Virology Journal recently, they explain that they constructed a RNAi lentiviral vector, iav pLenti6/BLOCK -iTTM, which expressed siRNA targeting the FMDV receptor, the porcine integrin av subunit, on PK-15 cells.

The researchers also produced a lentiviral stock, established an iav-PK-15 cell line, evaluated the gene silencing efficiency of mRNA using real-time qRT-PCR, integrand av 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.

Their results indicate the successful establishment of the iav U6 RNAi entry vector and the iav pLenti6/BLOCK -iT expression vector. The functional titre of obtained virus was 1.0 × 106TU/mL. To compare with the control and mock group, the iav-PK-15 group av mRNA expression rate in group was reduced by 89.5 per cent, whilst IIF and cell ELISA clearly indicated suppression in the experimental group. Thus, iav-PK-15 cells could reduce virus growth by more than three-fold and there was a more than 99 per cent reduction in virus titre when cells were challenged with 102 TCID50 of FMDV.

Luo and co-authors concluded that iav-PK-15 cells were demonstrated as a cell model for anti-FMDV potency testing and the study suggests that shRNA could be a viable therapeutic approach for controlling the severity of FMD infection and spread.

Reference

Luo J., J. Du, S. Gao, G. Zhang, J. Sun, G. Cong, J. Shao, T. Lin and H. Chang. 2011. Lentviral-mediated RNAi to inhibit target gene expression of the porcine integrin av subunit, the FMDV receptor, and against FMDV infection in PK-15 cells. Virology Journal 2011, 8:428. doi:10.1186/1743-422X-8-428

Further Reading

- You can view the full report by clicking here.


Further Reading

- Find out more information on foot and mouth disease (FMD) by clicking here.


November 2011