The hybridization chain reaction(HCR)is a widely used nucleic acid amplification technique that is essential for gene expression analysis and disease diagnosis.Despite its inherent stability,traditional HCR often suff...The hybridization chain reaction(HCR)is a widely used nucleic acid amplification technique that is essential for gene expression analysis and disease diagnosis.Despite its inherent stability,traditional HCR often suffers from low detection efficiency,which necessitates the use of supplementary molecular technologies to enhance its performance for detecting trace samples.Drawing insights from confinement theory,we have proposed a novel cruciform DNA scaffold-based HCR reaction system(C-HCR).In our approach,the DNA cruciform is assembled with the initiating probes of the HCR reaction through specific binding between adenine-rich and thymine-rich regions.This assembly with derived chains does not interfere with the HCR process.When a target is present,traditional HCR reactions yield long-chain linear products.However,the incorporation of the DNA cruciform in C-HCR leads to the formation of network-like products,which create favorable conditions for efficient molecular collisions and,in turn,promote high detection efficiency within the system.We utilized miR-21 and miR-27a as model targets to validate our design concept.Our results revealed that the miRNA-specific HCR system for miR-21 and miR-27a achieved significant increases in detection efficiency of 27.8%and 50%,respectively,demonstrating the feasibility and versatility of our design.This study offers a new strategy for enzyme-free amplification systems.展开更多
基金supported by the Natural Science Foundation of Anhui Province(2308085QH303)Anhui Provincial Health Research Program(AHWJ2022b113)Basic and Clinical Collaboration Program of Anhui Medical University(2022sfy004)。
文摘The hybridization chain reaction(HCR)is a widely used nucleic acid amplification technique that is essential for gene expression analysis and disease diagnosis.Despite its inherent stability,traditional HCR often suffers from low detection efficiency,which necessitates the use of supplementary molecular technologies to enhance its performance for detecting trace samples.Drawing insights from confinement theory,we have proposed a novel cruciform DNA scaffold-based HCR reaction system(C-HCR).In our approach,the DNA cruciform is assembled with the initiating probes of the HCR reaction through specific binding between adenine-rich and thymine-rich regions.This assembly with derived chains does not interfere with the HCR process.When a target is present,traditional HCR reactions yield long-chain linear products.However,the incorporation of the DNA cruciform in C-HCR leads to the formation of network-like products,which create favorable conditions for efficient molecular collisions and,in turn,promote high detection efficiency within the system.We utilized miR-21 and miR-27a as model targets to validate our design concept.Our results revealed that the miRNA-specific HCR system for miR-21 and miR-27a achieved significant increases in detection efficiency of 27.8%and 50%,respectively,demonstrating the feasibility and versatility of our design.This study offers a new strategy for enzyme-free amplification systems.
