Live-cell single RNA imaging reveals bursts of translational frameshifting

Kenneth Lyon; Luis U. Aguilera; Tatsuya Morisaki; Brian Munsky; Timothy J. Stasevich

The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https: //doi.org/10.1101/478040 doi: bioRxiv preprint To test this model, we co-transfected a short oligo RNA containing the FSS alone into cells expressing the +FSS MF tag. This led to a significant increase in the fraction of frameshifting sites translating just the -1 frame, from 1.6 to 5.6% when 1 g oligo was added, and up to 9.5% when 4 g oligo was added (Fig. 3A,B) . The significant increase in the -1 only sites suggests RNA can exist in a state in which all or nearly all ribosomes are frameshifted. Moreover, frameshifting continued to occur preferentially within factories, having RNA signals from 30 to 50% brighter than RNA-only spots (Fig. 3C) , even though the overall fraction of RNA within factories remained unchanged (Fig. S6 ).