A Deoxyribozyme-Initiated Self-Catalytic DNA Machine for Amplified Live-Cell Imaging of MicroRNA

Functional DNA nanostructures have been widely used in various bioassay fields. Yet, the programmable assembly of functional DNA nanostructures in living cells still represents a challenging goal for guaranteeing the sensitive and specific biosensing utility. In this work, we report a self-catalytic...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2021-08, Vol.93 (31), p.11052-11059
Main Authors: Wan, Yeqing, Li, Gaiping, Zou, Lina, Wang, Hong, Wang, Qing, Tan, Kaiyue, Liu, Xiaoqing, Wang, Fuan
Format: Article
Language:English
Subjects:
Amplification
Bioassays
Biomarkers
Biosensors
Catalysis
Cells (biology)
Chemistry
Deoxyribonucleic acid
DNA
Fluorescence
Medical imaging
MicroRNAs
miRNA
Modules
Nanostructure
Ribonucleic acid
RNA
Self-assembly
Substrates
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Summary:Functional DNA nanostructures have been widely used in various bioassay fields. Yet, the programmable assembly of functional DNA nanostructures in living cells still represents a challenging goal for guaranteeing the sensitive and specific biosensing utility. In this work, we report a self-catalytic DNA assembly (SDA) machine by using a feedback deoxyribozyme (DNAzyme)-amplified branched DNA assembly. This SDA system consists of catalytic self-assembly (CSA) and DNAzyme amplification modules for recognizing and amplifying the target analyte. The analyte initiates the CSA reaction, leading to the formation of Y-shaped DNA that carries two RNA-cleaving DNAzymes. One DNAzyme can then successively cleave the corresponding substrate and generate numerous additional inputs to activate new CSA reactions, thus realizing a self-catalytic amplification reaction. Simultaneously, the other DNAzyme is assembled as a versatile signal transducer for cleaving the fluorophore/quencher-modified substrate, leading to the generation of an amplified fluorescence readout. By incorporating a flexible auxiliary sensing module, the SDA system can be converted into a universal sensing platform for detecting cancerous biomarkers, e.g., a well-known oncogene microRNA-21 (miR-21). Moreover, the SDA system realized the precise intracellular miR-21 imaging in living cells, which is attributed to the reciprocal amplification property between CSA reactions and DNAzyme biocatalysis. This compact SDA amplifier machine provides a universal and facile toolbox for the highly efficient identification of cancerous biomarkers and thus holds great potential for early cancer diagnosis.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.1c02596