Stealth Fluorescence Labeling for Live Microscopy Imaging of mRNA Delivery.

ABSTRACT: Methods for tracking RNA inside living cells without perturbing their natural interactions and functions are critical within biology and, in particular, to facilitate studies of therapeutic RNA delivery. We present a stealth labeling approach that can efficiently, and with high fidelity, generate RNA transcripts, through enzymatic incorporation of the triphosphate of tC^O, a fluorescent tricyclic cytosine analogue. We demonstrate this by incorporation of tC^O in up to 100% of the natural cytosine positions of a 1.2 kb mRNA encoding for the histone H2B fused to GFP (H2B:GFP). Spectroscopic characterization of this mRNA shows that the incorporation rate of tC^O is similar to cytosine, which allows for efficient labeling and controlled tuning of labeling ratios f

SUBMITTER: Baladi T 

PROVIDER: S-EPMC8154517 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature