Using All Probes Plasmids to Lower the Barrier for Quantifying Single-mRNA Translation Dynamics in Live Cells

Translation of mRNA into full-length protein is a relatively short-lived and dynamic process. Many classical biochemical techniques lack the resolution required to capture both spatial and temporal information of ribosome activity-initiation, elongation, and termination. This limitation is exacerbated in a live cell context. Recent advances in plasmid expression vectors, epitope tags, and fluorescent proteins now allow for quantitative and stream-lined imaging of single-mRNA translation in living cells. This protocol describes the use of "All Probes Plasmids" to lower the barrier for imaging and quantifying single-mRNA translation events in living cells.