Sensing of biomolecular interactions using fluorescence complementing systems in living cells

Area of Science:

• Biochemistry
• Molecular Biology
• Cell Biology

Background:

• Understanding cellular mechanisms relies on sensing biomolecule interactions within living cells.
• Molecular biosensors, particularly fluorescence complementation techniques like BiFC and TriFC, are crucial for this sensing.
• These methods are vital for advancing clinical diagnostics.

Purpose of the Study:

• To review the principles and applications of bimolecular/trimolecular fluorescence complementation (BiFC/TriFC).
• To highlight recent advancements and challenges in fluorescence fragment complementation systems.
• To underscore the potential of these systems in research and clinical settings.

Main Methods:

• Utilizing bimolecular fluorescence complementation (BiFC) and trimolecular fluorescence complementation (TriFC).
• These methods rely on the re-establishment of fluorescence when non-fluorescent protein fragments are brought together by interacting biomolecules.
• The technique is noninvasive, sensitive, and does not require specialized laboratory equipment.

Main Results:

• BiFC and TriFC enable the detection of protein-protein interactions in living cells.
• Recent developments include multicolor BiFC for simultaneous detection of multiple interactions, RNA-protein interactions, and near-infrared systems for deep tissue imaging.
• The method has been successfully applied to various molecular interactions.

Conclusions:

• Fluorescence fragment complementation systems offer a simple, sensitive, and noninvasive approach to studying biomolecular interactions.
• Technological advancements are expanding the capabilities of BiFC/TriFC, including multicolor and deep-tissue imaging applications.
• These systems are poised for broad utilization in routine research and clinical diagnostics.