Mallanna Sunil Kumar, Rasool T J, Sahay Bikash, Aleyas Abi George, Ram Hira, Mondal Bimalendu, Nautiyal Binita, Premraj Avinash, Sreekumar E, Yadav M P
Biotechnology Laboratory, Indian Veterinary Research Institute, Mukteswar 263138, Nainital, Uttaranchal, India.
Virus Res. 2006 Feb;115(2):192-7. doi: 10.1016/j.virusres.2005.08.012. Epub 2005 Sep 30.
RNA interference (RNAi) mediated by double stranded small interfering RNA (siRNA) is a novel mechanism of post-transcriptional gene silencing. It is projected as a potential tool to inhibit viral replication. In the present paper, we demonstrate the suppression of replication of an avian herpes virus (Anatid Herpes Virus-1, AHV-1) by siRNA mediated gene silencing in avian cells. The UL-6 gene of AHV-1 that codes for a protein involved in viral packaging was targeted. Both cocktail and unique siRNAs were attempted to evaluate the inhibitory potential of AHV-1 replication in duck embryo fibroblast (DEF) cell line. DEF cells were chemically transfected with different siRNAs in separate experiments followed by viral infection. The observed reduction in virus replication was evaluated by cytopathic effect, viral titration and quantitative real time PCR (QRT-PCR). Among the three siRNA targets used the unique siRNA UL-B sequence was found to be more potent in antiviral activity than the cocktail and UL6-A-siRNA sequences.
由双链小干扰RNA(siRNA)介导的RNA干扰(RNAi)是一种转录后基因沉默的新机制。它被视为抑制病毒复制的潜在工具。在本文中,我们证明了在禽细胞中,通过siRNA介导的基因沉默可抑制禽疱疹病毒(鸭疱疹病毒1型,AHV-1)的复制。AHV-1的UL-6基因编码一种参与病毒包装的蛋白质,该基因被作为靶点。尝试使用混合siRNA和单一siRNA来评估AHV-1在鸭胚成纤维细胞(DEF)系中的复制抑制潜力。在单独的实验中,用不同的siRNA对DEF细胞进行化学转染,随后进行病毒感染。通过细胞病变效应、病毒滴定和定量实时PCR(QRT-PCR)评估观察到的病毒复制减少情况。在所使用的三个siRNA靶点中,发现单一siRNA UL-B序列在抗病毒活性方面比混合siRNA和UL6-A-siRNA序列更有效。