Novel fluorescent strategies for multiplexed detection of miRNAs by utilizing nucleic acid modifying-enzymes

Abstract

Over the past decade, microRNAs (miRNAs) have emerged as an invaluable biomarker that can improve the diagnosis and prognosis of cancer patients. Due to its clinical significance, considerable effort has been made to the development of novel strategies for the analysis of miRNA, but its small size and relatively low abundance, together with the sequence homology between same miRNA family members, make this a challenging task. In this thesis, we developed novel fluorescent strategies for multiplexed detection of miRNAs by utilizing nucleic acid modifying-enzymes. First, a multicomponent light-up sensor based on target-triggered transcription of RNA aptamers is developed for the one-step, multiplex sensing of microRNA. Second, a multiparameter suspension array platform utilizing splintR ligase-mediated ligation reaction and flow cytometry is developed for the multiplexed detection of miRNA. Next, a multicolor QD patterned array platform is developed based on DSN activity and highly uniform and precise patterning. Lastly, an enzyme-free, label-free strategy based on TMSD reaction is developed for the universal detection of miRNA.