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.
Narita
Atsushi
A
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan.
Ogawa
Kazumasa
K
Sando
Shinsuke
S
Aoyama
Yasuhiro
Y
D001483:Base Sequence; D015374:Biosensing Techniques; D002474:Cell-Free System; D003124:Colorimetry; D031541:Fluorescence Resonance Energy Transfer; D017930:Genes, Reporter; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D049408:Luminescent Agents / Q000032:analysis; D008163:Luminescent Measurements; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009696:Nucleic Acids / Q000032:analysis*; D009838:Oligodeoxyribonucleotides / Q000032:analysis; D014176:Protein Biosynthesis; D015347:RNA Probes / Q000737:chemistry*; D019713:Receptors, CCR5 / Q000235:genetics; D016914:Ribonuclease H; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D049408:Luminescent Agents; D009696:Nucleic Acids; D009838:Oligodeoxyribonucleotides; D015347:RNA Probes; D019713:Receptors, CCR5; D008156:Luciferases; D016914:Ribonuclease H; D001616:beta-Galactosidase