Single molecule viral RNA analysis using Stellaris™ FISH technology

Image courtesy of: http://www.cdc.gov/h1n1flu/images.htm

A cooperative effort between researchers at Mount Sinai School of Medicine in New York and the University of Illinois at Urbana-Champaign demonstrates a novel experimental system for single-molecule sensitivity FISH analysis of individual virus particles. 

Entitled “One influenza virus particle packages eight unique RNAs as shown by FISH analysis”, the paper describes the methods by which the investigators demonstrated that Influenza A viruses each package eight unique viral RNA segments, and provides a powerful tool to understand the stoichiometry & composition of segmented viral RNAs inside virus particles. One of the techniques used for the quantitation of the particles measures step-wise photobleaching characteristics of the fluorescent labels to determine copy numbers of RNA segments within a viral particle.

For our Stellaris FISH technology users, this article describes a very useful tool for:

• the study of viral packaging,
• a high throughput screening / drug discovery tool to test and/or confirm the disruption of the viral packaging process,
• studying the recombination of flu viruses during co-infection, and
• the study of other RNA viruses.

Yi-ying Chou, Reza Vafabakhsh, Sultan Doğanay, Qinshan Gao, Taekjip Ha, and Peter Palese.  One influenza virus particle packages eight unique RNAs as shown by FISH analysis.  Proc Natl Acad Sci USA 2012, Apr. 30. doi: 10.1073/pnas.1206069109.