A modular platform for bioluminescent RNA tracking
Ng, K. K.; Cole, K. H.; Halbers, L. H.; Chen, C.; Barhoosh, A. A.; Reid-McLaughlin, E.; Metcalfe, M.; Kent, A. D.; Steward, O.; Luptak, A.; Prescher, J. A.
Show abstract
A complete understanding of RNA biology requires methods for tracking transcripts in vivo. Common strategies rely on fluorogenic probes that are limited in sensitivity, dynamic range, and depth of interrogation, owing to their need for excitation light and tissue autofluorescence. To overcome these challenges, we developed a bioluminescent platform for serial imaging of RNAs. Small RNA tags were engineered to recruit light-emitting luciferase fragments (termed RNA lanterns) upon transcription. Robust photon production was observed for RNA targets both in cells and in live animals. Importantly, only a single copy of the tag was necessary for sensitive detection, in sharp contrast to fluorescent platforms requiring multiple repeats. Overall, this work provides a foundational platform for visualizing RNA dynamics from the micro to the macro scale.
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