Induction of protection against foot-and-mouth disease virus in cell culture and transgenic suckling mice by miRNA targeting integrin αv receptor

• Published in: Journal of biotechnology Vol. 187; pp. 154 - 161
• Main Authors: Du, Junzheng, Guo, Xinbing, Gao, Shandian, Luo, Jihuai, Gong, Xiuli, Hao, Chunxia, Yang, Bo, Lin, Tong, Shao, Junjun, Cong, Guozheng, Chang, Huiyun
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 10.10.2014
• Subjects: Animals; Animals, Suckling; Artificial miRNA; Biotechnology; Cell Line; Cricetinae; FMDV receptor; Foot-and-Mouth Disease - drug therapy; Foot-and-Mouth Disease - mortality; Foot-and-Mouth Disease Virus; Gene expression; Genes; Integrin alphaV - genetics; Integrin alphaV - metabolism; Mice; Mice, Transgenic; MicroRNAs - genetics; MicroRNAs - metabolism; MicroRNAs - therapeutic use; Receptors; Ribonucleic acids; RNAi; Survival Analysis; Swine; Transgenic; Transgenic suckling mouse; Virus infection; FMDV receptor; RNAi; Virus infection; Artificial miRNA; Transgenic suckling mouse
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2014.07.001

•The expression of three αv-specific miRNAs targeting integrin αv receptor inhibited FMDV infection in cell culture.•Transgenic mice were generated by the integration of the integrin αv-specific miRNA expression cassette into the mouse genome.•Transgenic suckling mice exhibited enhanced resistance against FMDV infection.•This is the first report of the use of miRNA-mediated gene knock down of host-cell receptors to inhibit FMDV infection in cell culture and transgenic suckling mice. Foot-and-mouth disease virus (FMDV) is an RNA virus that causes a highly contagious disease in domestic and wild cloven-hoofed animals. Although vaccination has been used to protect animals against FMDV, there are shortcomings in the efficacy of the available vaccines. RNA interference (RNAi) is triggered by small RNA molecules, including short interfering RNAs and microRNAs (miRNAs), and the use of RNAi-based methods have demonstrated promise as an alternative method of controlling the transmission of FMDV. However, the method of delivery, short duration of siRNA and miRNA in vivo, and the genetic variability of FMDV confound the use of RNAi-based strategies for FMDV control. FMDV has been shown to exploit host-cell integrins as cell-surface receptors to initiate infection. We selected the gene for the integrin αv subunit as an RNAi target, and constructed three αv-specific miRNA expression plasmids. The effects of these miRNAs on FMDV infection were examined in PK-15 cells and transgenic suckling mice. In PK-15 cells, the expression of the αv-specific miRNAs significantly inhibited the expression of integrin αv receptor and decreased FMDV infection. The transgenic mice were generated by integrating the αv-specific miRNA expression cassette using pronuclear microinjection. When challenged with a dose of FMDV ten times greater than the LD50, the survival rate of transgenic suckling mice was approximately six-fold higher than that of their non-transgenic littermates, indicating that the interference of the miRNAs significantly reduced FMDV infection in the transgenic mice. This is the first report of limiting FMDV attachment to cellular receptors using miRNA-mediated gene knock down of cell-surface receptors to significantly reduce FMDV infection in cell culture and transgenic suckling mice.