Cascaded and Localized Assembly of DNA Nanospheres for Efficient mRNA Imaging.
Liuting MoRunhong QinYan HongMingxiu MoChan YangWeiying LinPublished in: Analytical chemistry (2024)
The pursuit of advanced mRNA detection methods has been driven by the need for sensitive, accurate approaches that are particularly suited for live-cell analysis. Herein, we proposed a cascaded and localized assembly (CLA) system, integrating branched catalytic hairpin assembly (bCHA) with a localized hybridization chain reaction (LHCR) for enhanced mRNA imaging. The CLA system employed a dual-nanosphere (NS) platform, NSABC and NS12, and the interaction between the target and NSABC initiated the bCHA process and activated a split trigger. The newly generated trigger served as the initiator for the LHCR on NS12, leading to amplified fluorescent signals. Notably, this work introduced the first integration of a splitting strategy in a bCHA-HCR cascaded system, reducing false-positive signals and enhancing specific detection. The dual-NS platform further minimized background noise and improved the reaction kinetics through spatial confinement. As a result, the system achieved a detection limit of 1.23 pM. With these advantages, the CLA system demonstrated successful application in both living cells and clinical tissues, underscoring its potential in biomolecular research and clinical diagnostics.
Keyphrases
- living cells
- label free
- dengue virus
- single molecule
- high resolution
- loop mediated isothermal amplification
- fluorescent probe
- air pollution
- real time pcr
- gene expression
- particulate matter
- quantum dots
- zika virus
- cell free
- heavy metals
- photodynamic therapy
- mass spectrometry
- circulating tumor
- polycyclic aromatic hydrocarbons
- data analysis
- circulating tumor cells