Multicolor-Encoded DNA Framework Enables Specific and Amplified In Situ Detection of the Mitochondrial Apoptotic Signaling Pathway

Monitoring the molecular activation networks of cellular processes through fluorescence imaging to accurately elucidate the signaling pathways of mitochondrial apoptosis and the regulation of upstream and downstream molecules remains a current major challenge. In this work, a multicolor-encoded tetr...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2023-08, Vol.95 (33), p.12514-12520
Main Authors: Yang, Fang, Li, Shunmei, Bi, Xin, Yuan, Ruo, Xiang, Yun
Format: Article
Language:English
Subjects:
Amplification
Analytical chemistry
Apoptosis
Biocompatibility
Cellular communication
Chemical synthesis
Cytochrome c
Cytochromes
Deoxyribonucleic acid
Detection limits
DNA
DNA probes
Drug screening
Drugs
Fluorescence
Intracellular
Manganese
Mitochondria
mRNA
Reaction kinetics
Signal monitoring
Signal transduction
Superoxide dismutase
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Summary:Monitoring the molecular activation networks of cellular processes through fluorescence imaging to accurately elucidate the signaling pathways of mitochondrial apoptosis and the regulation of upstream and downstream molecules remains a current major challenge. In this work, a multicolor-encoded tetrahedral DNA framework (meTDF) carrying two pairs of catalytic hairpins is synthesized to monitor the intracellular upstream manganese superoxide dismutase (MnSOD) mRNA and the downstream cytochrome c (Cyt c) molecules for specific and sensitive detection of the mitochondrial apoptotic signaling pathway. These two types of molecules can trigger catalytic hairpin assembly (CHA) reactions with accelerated reaction kinetics for the hairpin pairs confined on meTDF to show highly amplified fluorescence for sensitive and simultaneous detection of MnSOD mRNA and Cyt c with detection limits of 3.7 pM and 0.23 nM in vitro, respectively. Moreover, the high stability and biocompatibility of the designed meTDF can facilitate efficient delivery of the probes into cells to monitor intracellular MnSOD mRNA and Cyt c for specific detection of the mitochondrial apoptosis pathway regulated by different drugs. With the successful demonstration of their robust capability, the meTDF nanoprobes can thus open new opportunities for detecting cell apoptotic mechanisms for studying the corresponding apoptotic signaling pathways and for screening potential therapeutic drugs.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.3c02462