shRNA表达质粒体外防治猪传染性胃肠炎病毒感染

shRNA-expressing Plasmids Prevent Transmissible Gastroenteritis Virus Infection in vitro

猪传染性胃肠炎病毒是引起猪胃肠道感染的主要病原之一,新生仔猪感染后死亡率很高,给养猪业带来重大经济损失。为了探索运用RNA干扰技术防治猪传染性胃肠炎病毒感染的可能性,我们构建了两个靶向病毒基因组的小发卡RNA (shRNA)表达质粒pEGFP-U6/P1和pEGFP-U6/P2,并分别转染猪睾丸(ST)细胞。通过细胞增殖试验(MTS)和定量RT-PCR检测试验表明,这两个质粒均可高效特异地抑制猪传染性胃肠炎病毒的复制。研究结果表明,RNA干扰(RNAi)技术可以用来有效防治猪传染性胃肠炎。

Porcine transmissible gastroenteritis virus （TGEV） is identified as one of the major pathogenic agents during swine entenc infection, leading to high mortality in neonatal pigs and severe annual economic loss in swine-producing areas. To investigate the possibility of using RNA interference （RNAi） as a strategy for anti-TGEV infection, two shRNA-expressing plasmids （pEGFP-U6/P1 and pEGFP-U6/P2） targeting TGEV genome were constructed and transfected into swine testicular （ST） cells. MTS assay and realtime quantitative RT-PCR both demonstrated that the two shRNA-expressing plasmids were capable of protecting ST cells from TGEV destruction with very high specificity and efficiency. Taken together, our results suggest that RNAi might become a promising new strategy for anti-TGEV infection.