SunRISE: long-term imaging of individual mRNA molecules in living cells

Abstract

Single-cell imaging of individual mRNAs has revealed core mechanisms of the central dogma. However, most approaches require cell fixation or have limited sensitivity for live-cell applications. Here, we describe SunRISE (SunTag-based Reporter for Imaging Signal Enriched mRNA), a computationally and experimentally optimized approach for unambiguous single-mRNA detection in living cells. We demonstrate SunRISE with long-term epifluorescence imaging, using translational stress to track mRNA phase separation and recovery from cytosolic droplets.