Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies

STAR Protoc. 2023 Mar 17;4(1):102033. doi: 10.1016/j.xpro.2022.102033. Epub 2023 Jan 14.

Abstract

Characterization of double-stranded (ds)RNAs is relevant to the understanding of viral replication and immune sensing. Here, we provide a protocol describing the use of anti-dsRNA antibodies for immunofluorescence and immunoblotting in virus-infected insect cells, which can also be applied to tissues and other organisms. We describe the procedures to prepare insect cells for viral infection, followed by RNA extraction and in vitro production of synthetic dsRNA controls. We then detail the steps for dsRNA detection by immunoblotting and immunofluorescence. For complete details on the use and execution of this protocol, please refer to de Faria et al. (2022).1.

Keywords: Antibody; Cell Biology; Immunology; Microbiology; Microscopy; Molecular Biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescent Antibody Technique
  • Immunoblotting
  • Insect Viruses* / genetics
  • Insecta* / cytology
  • Insecta* / virology
  • RNA, Double-Stranded*

Substances

  • RNA, Double-Stranded