Nucleic acid-based fluorescent probes and their analytical potential

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Nucleic acids are vital for genetic information storage and protein synthesis.
• The ligand-binding capabilities of nucleic acids and their use as molecular probes/biosensors are less understood.
• Oligonucleotide probes, single-stranded nucleic acid fragments, exhibit high specificity and affinity for diverse targets.

Purpose of the Study:

• To review the fundamentals of designing and engineering oligonucleotide probes.
• To describe typical construction approaches for these probes.
• To discuss examples of probes used in hybridization and aptamer-based assays.

Main Methods:

• Categorization of oligonucleotide probes into hybridization and aptamer types.
• Modification of oligonucleotide sequences with reporter groups, typically fluorophores.
• Utilizing fluorescent labels as transducers to convert biorecognition events into fluorescence signals.

Main Results:

• Oligonucleotide probes can be designed for high specificity and affinity towards nucleic acids, proteins, small molecules, and ions.
• Fluorescent labels offer advantages like high sensitivity and multiple transduction approaches (e.g., FRET, fluorescence anisotropy).
• The flexibility in probe design (DNA, RNA, PNA, LNA) and labeling strategies enables the development of selective and sensitive bioassays.

Conclusions:

• Oligonucleotide probes are versatile tools for molecular detection and bioassays.
• Advancements in design, construction, and labeling strategies enhance probe sensitivity and selectivity.
• These probes have broad applications in hybridization studies and aptamer-based assays.