With Influenza A virus, regardless of antigenic drift or shift we see reassortment of the 8 RNP segments. Is there a VLP or recombinant influenza strain that has 8 individually fluorescent labelled RNPs? Thus allowing further studies in re- assortment dynamics on mixed populations in vivo and in ex vivo cell lines?
Sadly no, at least not that I know of. And that's whether you mean actually labelling the individual RNPs, or the gene products the 8 segments encode. FISH can be used to stain fixed cells for individual segments, but your wording makes me suspect you want live cell methods? The Fodor lab has figured out how to put stem-loop structures into a segment that can be picked up by a sequence-specific GFP-RNA binding domain fusion, but how many options there are for that, I don't know. In terms of labelling individual virus proteins with FPs, yes this can be done (Google "colorflu" or similar) but I would be cautious about using them for a reassortant study because flu doesn't like have a large gene inserted into it (terminal packaging signals are a particular problem) and many if not all of the GFP/RFP/Luc/Etc-tagged viruses have fitness penalties compared to WT.
Cheers
Paul
Many thanks Paul,
yes I did fear that insertion of any type of marker FP would prove problematic and be booted out pretty early. I've used m-cherry with alphavirus infectious clones in the past which hasn't appeared to affect fitness, but flu is a different beast altogether. Many thanks for the insights here, I will follow up on the suggestions you've made and see what live cell methods I can find.
Best wishes
Christopher
That is such an interesting idea! I hope you succeed doing that Christopher! Influenza, but on the vaccine development side, is one of my focus areas. Best wishes for success!
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