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The copyright holder for this preprint (which was not peer-reviewed) is the author/funder.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 sites translating just the -1 frame (1.6 ± 0.5%, Fig. S2 and Movie S4). To ensure these results were not influenced by the multi-frame tag, we reversed the FLAG and SunTag epitopes in the tag and repeated experiments. This gave nearly the same percentages, confirming the tag order and/or epitope positioning did not bias measurements ( Fig. S3 and Movie S5). We then repeated experiments in cells transfected with the control -FSS construct. In this case, we observed virtually no frameshifting sites (0.9 ± 0.7%) ( Fig. 2C and Movie S6). Taken together, these data suggest the FSS alone causes ~8% of translation sites to frameshift.