慢病毒介导siRNA抑制O型口蹄疫病毒ON株病毒复制

doi:10.11843/j.issn.0366-6964.2018.02.015

Abstract

Abstract:

In order to inhibit replication of foot-and-mouth disease virus (FMDV) by RNAi technology, two pairs of short hairpin RNA (shRNA) targeting 3D, 3C and VP1 gene of foot-and-mouth disease virus ON strain were designed separately and named as VP1-1,VP1-2, 3C-1, 3C-2, 3D-1 and 3D-2. According to the principle of RNAi, the six shRNA expression plasmid were constructed and transfected into BHK21 cell that was infected with the FMDV ON strain. TCID₅₀ and real-time fluorescent quantitative PCR were used to analyze inhibition efficiency of shRNA recombinant plasmids on FMDV. ShRNAs with highest inhibition efficiency were chose to produce lentivirus particles and infect BHK21 cell lines, TCID₅₀ and real-time fluorescent quantitative PCR were used to analyze lentivirus inference vectors inhibition efficiency on foot-and-mouth disease virus. The results showed that the inhibition rate of shRNAs was between 71.5%-93.2%, and the most obvious inhibitory effect were PLKO.1-VP1-2 as 93.2% and PLKO.1-3D as 90.8%, respectively; lentivirus inference vectors PLKO.1-VP1-2 and PLKO.1-3D inhibition rate of FMDV were between 88.3%-95.49%. Our result screened out shRNAs that can inhibit foot-and-mouth disease virus replication at the cellular level. The study laid the foundation for transgenic sheep against FMDV production and cultivation.

CLC Number:

S852.659.6

QI Xing-cai, QIN Xiao-dong, GAN Xiao-li, ZHANG Shu-min, MA You-ji, LI Zhi-yong. Inhibition of Type O Foot-and-mouth Disease Virus ON Strain Replication by Lentivirus-mediated siRNA[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(2): 360-367.

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Inhibition of Type O Foot-and-mouth Disease Virus ON Strain Replication by Lentivirus-mediated siRNA

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