Visible sensing of nucleic acid sequences with a genetically encodable unmodified RNA probe.
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Base Sequence↗
Biomedical subjects
Kazumasa Ogawa
Publications and source records attributed to Kazumasa Ogawa.
Visible sensing of nucleic acid sequences using a genetically encodable unmodified mRNA probe.
We previously reported a molecular beacon-mRNA (MB-mRNA) strategy for nucleic acid detection/sensing in a cell-free translation system using unmodified RNA as a probe. Here in this presentation, we report that a combination with RNase H activity, which induces an additional process of irreversible cleavage of MB-domain, achieves an improved sequence selectivity (one nucleotide selectivity) and an enhanced sensitivity. This improved system finally enabled visible sensing of target nucleic acid sequence at a single nucleotide resolution under isothermal conditions.
Base Sequence↗
Highly sensitive genotyping using artificial riboregulator system.
A molecular-beacon-type riboregulator (mRNA) was applied to multiply catalytic gene sensing. It consists of a reporter gene for firefly protein luciferase and, upstream thereof, a regulator hairpin domain composed of an RBS/anti-RBS stem (RBS = ribosome binding site) and a loop which is complementary to the target. The sensing of target gene, using an unmodified RNA or even dsDNA as a probe with a chemiluminescence output, was demonstrated with a sensitivity at < or = 50 fmol of the target and a single nucleotide resolution.
Bacterial Proteins↗