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A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons
Published on: November 14, 2018
Size-Controllable and Self-Assembled DNA Nanosphere for Amplified MicroRNA Imaging through ATP-Fueled Cyclic
Jiaoli Wang1, Juan Li1, Yu Chen1
1State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082, P.R. China.
Abstract:
Visualizing intracellular microRNA (miRNA) is of great importance for revealing its roles in the development of disease. However, cell membrane barrier, complex intracellular environment and low abundance of target miRNA are three main challenges for efficient imaging of intracellular miRNA. Here, we report a size-controllable and self-assembled DNA nanosphere with ATP-fueled dissociation property for amplified miRNA imaging in live cells and mice. The DNA nanosphere was self-assembled from Y-shaped DNA (Y-DNA) monomers through predesigned base pair hybridization, and the size could be easily controlled by varying the concentration of Y-DNA. Once the nanosphere was internalized into cells, the intracellular specific target miRNA would trigger the cyclic dissociation of the DNA nanosphere driven by ATP, resulting in amplified FRET signal. The programmable DNA nanosphere has been proven to work well for detecting the expression of miRNA in cancer cells and in mice, which demonstrates its fairish cell penetration, stability and sensitivity.
Insights
Researchers developed a novel DNA nanosphere for enhanced intracellular microRNA (miRNA) imaging. This ATP-fueled system offers improved detection in live cells and mice, overcoming key imaging challenges.
Area of Science:
- Biomedical Engineering
- Molecular Biology
- Nanotechnology
Background:
- Visualizing intracellular microRNA (miRNA) is crucial for understanding disease mechanisms.
- Challenges include the cell membrane barrier, complex intracellular environments, and low miRNA abundance.
- Existing methods struggle with efficient and sensitive intracellular miRNA detection.
Purpose of the Study:
- To develop a size-controllable, self-assembled DNA nanosphere for amplified intracellular miRNA imaging.
- To overcome the limitations of current miRNA detection methods in live cells and organisms.
- To demonstrate the efficacy of the DNA nanosphere system in detecting miRNA expression in cancer models.
Main Methods:
- Self-assembly of Y-shaped DNA (Y-DNA) monomers into size-controllable DNA nanospheres via base-pair hybridization.
- Utilizing ATP-fueled cyclic dissociation of the nanosphere triggered by intracellular target miRNA.
- Employing Förster Resonance Energy Transfer (FRET) for signal amplification.
- Testing the system in live cells and live mice models.
Main Results:
- The DNA nanosphere system demonstrated size controllability by adjusting Y-DNA concentration.
- Intracellular target miRNA triggered cyclic nanosphere dissociation, leading to amplified FRET signals.
- The system showed effective miRNA detection in cancer cells and in mice.
- The nanosphere exhibited good cell penetration, stability, and sensitivity.
Conclusions:
- The developed DNA nanosphere is a promising tool for amplified intracellular miRNA imaging.
- The ATP-fueled dissociation mechanism enhances detection sensitivity.
- This technology holds potential for disease diagnosis and research, particularly in cancer studies.
