Detection Of MicroRNA Based On Rolling Circle Amplification And Fluorescence Resonance Energy Transfer Triggered By The Front End

Detection of microRNAs(miRNAs)in cells can help improve our understanding of the physiological functions of miRNAs and can facilitate the exploration of their relationships with disease.The toehold-mediated strand displacement reaction initiates rolling circle amplification(RCA)to achieve signal amplification of the miRNA to be tested;this process is called toehold-initiated RCA(TIRCA).The product of TIRCA was ligated to two DNA probes,each modified with 6-carboxyfluorescein and carboxytetramethylrhodamine.Qualitative detection of miRNAs was successfully achieved by combining the fluorescence aggregation enhancement effect with fluorescence resonance energy transfer generated by the proximity of the two fluorescent dyes.Our results showed that this approach can help us analyze the roles of miRNAs in human disease more accurately.The third chapter introduces the preparation of miR-494 and miR-6126 in different cells to explore their physiological functions.The probe for detection of miR-6126 was selected to verify successful synthesis of the probe and formation of the reaction product.This lays the foundation for further exploration of the physiological functions of miRNAs.