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Updated: Mar 20, 2026

Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
Published on: August 6, 2014
Protocol to measure transcriptional bursting of endogenous genes using high-throughput RNA-FISH
1National Cancer Institute, Bethesda, MD 20892, USA.
Abstract:
We present a protocol to measure transcriptional bursting of endogenous human genes that is widely applicable to adherent cells. We describe the steps for nascent RNA fluorescence in situ hybridization (nscRNA-FISH) in a high-throughput imaging format. We then detail the procedures for automated microscopy and analysis to measure bursting behavior of genes in adherent cell lines. The frequency of bursting alleles in a population is used as a readout for bursting behavior. For complete details on the use and execution of this protocol, please refer to Sood et al.1.
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