Chen Mo, Payne William S, Hunt Henry, Zhang Huanmin, Holmen Sheri L, Dodgson Jerry B
Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI 48824, USA.
Virology. 2008 Aug 1;377(2):265-72. doi: 10.1016/j.virol.2008.03.019.
RNA interference (RNAi) is a promising antiviral methodology. We recently demonstrated that retroviral vectors expressing short-hairpin RNAs (shRNA-mirs) in the context of a modified endogenous micro-RNA (miRNA) can be effective in reducing replication of other retroviruses in chicken cells. In this study, similar RNAi vectors are shown to inhibit replication of the avian herpesvirus, Marek's disease virus (MDV, also known as gallid herpesvirus type 2), and its close relative, herpesvirus of turkeys (HVT). Cells expressing shRNA-mirs targeting the MDV or HVT gB glycoprotein gene or the ICP4 transcriptional regulatory gene show significant inhibition of viral replication. Not only are viral titers reduced, but observed plaque sizes are significantly smaller when the virus is grown on cells in which RNAi is effective. We also describe a modified retroviral delivery vector that expresses a shRNA-mir containing up to three RNAi target sequences and employ this vector with multiple targets within the MDV gB gene or within both the gB and ICP4 genes. The use of targets within multiple genes potentially can provide a larger antiviral effect and/or make it more difficult for viral escape mutations to evolve.
RNA干扰（RNAi）是一种很有前景的抗病毒方法。我们最近证明，在修饰的内源性微小RNA（miRNA）背景下表达短发夹RNA（shRNA-mirs）的逆转录病毒载体可有效减少鸡细胞中其他逆转录病毒的复制。在本研究中，类似的RNAi载体显示可抑制禽疱疹病毒、马立克氏病病毒（MDV，也称为2型鸡疱疹病毒）及其近亲火鸡疱疹病毒（HVT）的复制。表达靶向MDV或HVT gB糖蛋白基因或ICP4转录调节基因的shRNA-mirs的细胞显示出对病毒复制的显著抑制。不仅病毒滴度降低，而且当病毒在RNAi有效的细胞上生长时，观察到的噬斑大小也显著变小。我们还描述了一种修饰的逆转录病毒递送载体，其表达包含多达三个RNAi靶序列的shRNA-mir，并将该载体用于MDV gB基因内或gB和ICP4基因内的多个靶标。在多个基因内使用靶标可能会提供更大的抗病毒效果和/或使病毒逃逸突变更难发生。
