研究目的
Developing a visual detection method for miRNA 155 based on DNAzyme amplification strategy in living cells for early diagnosis of cancer.
研究成果
The developed method offers a highly sensitive and selective detection of miRNA 155 in living cells with a wide linearity range and low detection limit, contributing to tumor cells tracking and clinical definite in early-stage cancer.
研究不足
The method requires optimization of the concentrations of AuNPs-Probe and DNA modules for effective cell imaging. The incubating time for AuNPs-Probe and DNA modules with cells needs to be carefully controlled to maintain cell activity.
1：Experimental Design and Method Selection:
A DNAzyme-based amplification strategy was designed for the detection of miRNA 155 both in vitro and in living cells. MnO2 nanosheets were used to deliver DNA modules into cells, and AuNPs-Probe were used for fluorescence imaging.
2：Sample Selection and Data Sources:
HepG2 cells and LO2 cells were used as target and control cells for miRNA 155 imaging.
3：List of Experimental Equipment and Materials:
AuNPs, MnO2 nanosheets, DNA modules (DNAzyme, Locker DNA, Substrate DNA, FL DNA), and confocal laser microscopy were used.
4：Experimental Procedures and Operational Workflow:
DNA modules were delivered into cells using MnO2 nanosheets. MiRNA 155 activated the DNAzyme, which cleaved substrate DNA to release Key DNA. Key DNA opened hairpin DNA on AuNPs to turn on fluorescence.
5：Data Analysis Methods:
Fluorescence intensity was measured to quantify miRNA 155 levels.
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