Light-activatable molecular beacons with a caged loop sequence.

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Molecular beacons are DNA/RNA probes used for nucleic acid detection.
• Existing methods for RNA tracking in live cells face limitations in spatial and temporal resolution.
• Controlling probe fluorescence activation is crucial for precise molecular imaging.

Purpose of the Study:

• To synthesize and characterize novel molecular beacons with light-activatable caged nucleobases.
• To demonstrate the light-induced fluorescence activation of these probes in vitro and in live cells.
• To explore the potential of this technology for single-molecule RNA tracking in living systems.

Main Methods:

• Synthesis of molecular beacons incorporating caged nucleobases in the loop region.
• In vitro characterization of probe fluorescence in the presence and absence of target RNA.
• Testing probe activation and performance in HEK293 cells using specific light wavelengths (366 nm and 405 nm).

Main Results:

• The synthesized molecular beacons remained non-fluorescent when bound to target RNA.
• Light activation (366 or 405 nm) successfully triggered full fluorescence of the probes.
• The probes demonstrated light-inducible fluorescence activation both in vitro and within living HEK293 cells.

Conclusions:

• Molecular beacons with caged nucleobases offer a novel approach for light-controlled RNA detection.
• This technology provides a mechanism for activating fluorescence only upon illumination, reducing background noise.
• The developed probes show promise for advanced applications like single-molecule RNA tracking in live cells.