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miRNA Cell Tracer: Multifunctional Microgels for Spatially Resolved and Wide-Range Detection of Intracellular miRNA
Sabrina Napoletano1,2, Maria Isabella Maremonti1,2, Edmondo Battista3
1Interdisciplinary Research Centre on Biomaterials (CRIB), University of Naples "Federico II", Naples 80125, Italy.
ACS Sensors
|February 10, 2026
Summary
This study introduces microgels for sensitive, amplification-free detection of intracellular microRNAs (miRNAs) in live cells. The novel biosensor allows for spatial mapping and quantification of miRNAs, aiding gene regulation studies.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- Intracellular microRNA (miRNA) detection is crucial for understanding gene regulation.
- Existing methods face challenges with low miRNA abundance, heterogeneity, and intracellular delivery.
- Particle biosensors for live-cell miRNA detection are still limited.
Purpose of the Study:
- To develop an amplification-free method for sensitive and spatially resolved detection of intracellular miRNAs in live cells.
- To create a biosensor capable of delivering probes into the cytosol without affecting cell viability or endogenous miRNA levels.
- To enable quantitative monitoring of miRNA expression and localization at the single-cell level.
Main Methods:
- Utilized large (∼600 nm) biocompatible microgels functionalized with molecular beacons.
- Employed microfluidic mechanoporation for efficient microgel delivery into the cytosol.
- Developed an amplification-free detection system for target miRNA hybridization and fluorescence signal generation.
Main Results:
- Achieved a low-picomolar limit of detection (5.6 pM) and a wide dynamic range (pM-nM) for intracellular miRNA sensing.
- Demonstrated excellent specificity against nontarget and precursor miRNAs.
- Successfully quantified endogenous miRNAs (miR-191-5p, miR-363-5p) in healthy and cancer breast cells, revealing cell-to-cell variability and distinct localization patterns.
Conclusions:
- The microgel-based biosensor enables sensitive, quantitative, and spatially resolved amplification-free detection of intracellular miRNAs in live cells.
- This versatile platform overcomes limitations of conventional methods for intracellular miRNA analysis.
- The technology holds potential for live-cell diagnostics, therapeutic profiling, and single-cell gene regulation studies.
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