Primary Selection of MicroRNA Acting on Classical Swine Fever Virus in Swine Umbilical Vein Endothelial Cells

The aim of the study was to identify the microRNAs that act on classical swine fever virus (CSFV) in swine umbilical vein endothelial cells (SUVECs). The 3'-and 5'-untranslated regions ( UTR) of CSFV were cloned and then inserted into psiCHECH TM-2 plasmids carrying Firefly and Renilla luciferases reporter genes; microRNAs that acted with CSFV were predicted by bio-informatics analysis; then the recombinant plasmids and inhibitors of the predicted microRNA were co-transfected into SUVECs. The activities of Firefly and Renilla luciferases were detected by luminometry. The PCR products of the CSFV 5'-UTR (373 bp) and 3'-UTR (252 bp) were detected by electrophoresis on 1% agarose gels. Four microRNAs (ssc-miR-let7c, ssc-miR-106a, ssc-miR-18, ssc-miR-139) were screened out and evaluated. The CSFV 3'-UTR is the important target site of microRNA in SUVECs. The four microRNAs mentioned above had different inhibitory effects on the CSFV 3'-UTR, of which the ssc-miR-18 played the most important role.

The aim of the study was to identify the microRNAs that act on classical swine fever virus （CSFV） in swine umbilical vein endothelial cells （SUVECs）. The 3＇- and Y-untranslated regions （UTR） of CSFV were cloned and then inserted into psiCHECHTM-2 plasmids carrying Firefly and Renilla luciferases reporter genes; microRNAs that acted with CSFV were predicted by bio-informatics analysis; then the recombinant plasmids and inhibitors of the predicted microRNA were co-transfected into SUVECs. The activities of Firefly and Renilla luciferases were detected by luminometry. The PCR products of the CSFV 5＇-UTR （373 bp） and 3＇-UTR （252 bp） were detected by electrophoresis on 1% agarose gels. Four microRNAs （ssc-miR-let7c, ssc-miR-106a, ssc-miR-18, ssc-miR- 139） were screened out and evaluated. The CSFV 3＇-UTR is the important target site of microRNA in SUVECs. The four microRNAs mentioned above had different inhibitory effects on the CSFV 3＇-UTR, of which the ssc-miR-18 played the most important role.