通过果蝇S2细胞培养中的报告基因检测鉴定RNA干扰的病毒抑制因子。

Identification of viral suppressors of RNAi by a reporter assay in Drosophila S2 cell culture.

作者信息

van Cleef Koen W R, van Mierlo Joël T, van den Beek Marius, van Rij Ronald P

机构信息

Department of Medical Microbiology, Radboud University Nijmegen Medical Centre, Nijmegen Centre for Molecular Life Sciences, Nijmegen Institute for Infection, Inflammation and Immunity, Nijmegen, The Netherlands.

出版信息

Methods Mol Biol. 2011;721:201-13. doi: 10.1007/978-1-61779-037-9_12.

DOI:10.1007/978-1-61779-037-9_12

PMID:21431687

Abstract

The RNA interference (RNAi) pathway plays an important role in antiviral immunity in insects. To -counteract the RNAi-mediated immune response of their hosts, several insect viruses, such as Flock house virus, Drosophila C virus, and Cricket paralysis virus, encode potent viral suppressors of RNAi (VSRs). Because of the importance of RNAi in antiviral defense in insects, other insect viruses are likely to encode VSRs as well. In this chapter, we describe a detailed protocol for an RNAi reporter assay in Drosophila S2 cells for the identification of VSR activity.

摘要

RNA干扰（RNAi）途径在昆虫的抗病毒免疫中发挥着重要作用。为了对抗宿主的RNAi介导的免疫反应，几种昆虫病毒，如禽舍病毒、果蝇C病毒和蟋蟀麻痹病毒，编码了强大的RNAi病毒抑制因子（VSRs）。由于RNAi在昆虫抗病毒防御中的重要性，其他昆虫病毒也可能编码VSRs。在本章中，我们描述了一种在果蝇S2细胞中进行RNAi报告基因检测以鉴定VSR活性的详细方案。