Cells have developed various means for transporting molecules across membranes as well as cellular compartments separated by membranes. Channels and pores can be formed with different protein assemblies to enable the regulated passage of molecules in and out of cells, which has led to them being exploited as molecular sensors for characterizing biomolecules and developing assays for use as biological tools and for various biomedical applications. Nanopore technologies have been widely explored for their potential in biosensing and diagnostic applications, but most existing technologies employ in vitro demonstrations and membrane disruptive approaches.
Researchers at the Biodesign Institute of Arizona State University have developed a novel DNA hairpin-based nanopore capable of selective opening in response to binding nucleic acid targets, such as mRNA or miRNA. The nanopore amplifies signals by allowing extracellular fluorescent molecules to enter the cell after binding to the target, thus enabling sensitive detection of specific nucleic acid sequences without disrupting the cell. The reversible nature of the system provides further control, making it an ideal platform for molecular sensing and signal amplification. This nanopore has been tested with small molecule (e.g., Alexa fluorophores) and short peptide (e.g., phalloidin) as delivery cargo with Cy3 and Cy5 labeling reporters.
This new DNA hairpin nanopore enables lysis-free sensing so as to maintain cell integrity and allow for real-time monitoring of nucleic acid targets.
Potential Applications
- Gated Nanopore, which can be opened by binding to biomarkers
- Nucleic acid sensing – genomics, sequencing, analyte detection, etc.
- Signal amplification
- Delivery
- Molecular diagnostics – specifically nucleic acid sensing within living cells
- Live-cell genotyping and isolations
- Continuous monitoring of biomarkers in wearable devices
Benefits and Advantages
- Able to sense intracellular nucleic acids without lysing the cells, allowing for real-time monitoring and preserving cell function
- The signal amplification mechanism ensures a highly sensitive detection system, where a single binding event leads to the influx of multiple fluorophores
- The reversible nature of the nanopore allows for greater flexibility and control in experimental setups, enhancing its utility in biosensing applications
- The non-destructive sensing and signal amplification is compatible with live-cell isolations
- Fast signal amplification kinetics enabled by the nanopore's structural design
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