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Updated: Jan 31, 2026

Measuring the Kinetics of mRNA Transcription in Single Living Cells
Published on: August 25, 2011
ShortCut RNase III is capable of digesting single mRNA transcripts into < 10 nucleotide products
Kyle J Tynan1, Mark S Lowenthal2, Karndeep Singh3
1National Institute of Standards and Technology (NIST), 100 Bureau Dr, Gaithersburg, MD 20899 USA; Institute for Bioscience and Biotechnology Research (IBBR), 9600 Gudelsky Dr, Rockville, MD 20850 USA.
None:
RNase III is a dsRNA-specific ribonuclease that has been used to enzymatically digest dsRNA impurities generated during in vitro transcription. Here, we demonstrate that ShortCut RNase III, a recombinant version of E. coli RNase III fused with maltose binding protein which is reported to digest dsRNA to 18-25 base pair fragments, can also degrade single-stranded Cas9 mRNA to fragments as short as 9 nucleotides. These findings indicate that ShortCut RNase III exhibits unexpected activity beyond canonical dsRNA cleavage, raising concerns about its suitability for removing dsRNA impurities in mRNA manufacturing.
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