Orsay Virus Infection in Caenorhabditis elegans

Abstract

Orsay virus infection in the nematode Caenorhabditis elegans presents an opportunity to study host-virus interactions in an easily culturable, whole-animal host. Previously, a major limitation of C. elegans as a model for studying antiviral immunity was the lack of viruses known to naturally infect the worm. With the 2011 discovery of the Orsay virus, a naturally occurring viral pathogen, C. elegans has emerged as a compelling model for research on antiviral defense. From the perspective of the host, the genetic tractability of C. elegans enables mechanistic studies of antiviral immunity while the transparency of this animal allows for the observation of subcellular processes in vivo. Preparing infective virus filtrate and performing infections can be achieved with relative ease in a laboratory setting. Moreover, several tools are available to measure the outcome of infection. Here, we describe workflows for generating infective virus filtrate, achieving reproducible infection of C. elegans, and assessing the outcome of viral infection using molecular biology approaches and immunofluorescence. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC.

Basic Protocol 1: Preparation of Orsay virus filtrate

Support Protocol: Synchronize C. elegans development by bleaching

Basic Protocol 2: Orsay virus infection

Basic Protocol 3: Quantification of Orsay virus RNA1/RNA2 transcript levels by qRT-PCR

Basic Protocol 4: Quantification of infection rate and fluorescence in situ hybridization (FISH) fluorescence intensity

Basic Protocol 5: Immunofluorescent labeling of dsRNA in virus-infected intestinal tissue