Genetically-Encoded Phase Separation Sensors Enable High-Fidelity Live-Cell Probing of Biomolecular Condensates

Area of Science:

• Cell Biology
• Biophysics

Background:

• Biomolecular condensates are membraneless compartments crucial for cellular functions.
• Intrinsically disordered proteins (IDPs) often scaffold these condensates via liquid-liquid phase separation (LLPS).
• Current methods using fluorescently tagged IDPs are prone to artifacts due to protein conformational heterogeneity.

Purpose of the Study:

• To develop and optimize genetically-encoded sensors for probing epidermal biomolecular condensates.
• To assess the impact of these sensors in early and late stages of intracellular phase separation.
• To provide a high-fidelity, artifact-free method for studying IDP-governed condensates.

Main Methods:

• Development of tunable epidermal LLPS-sensors.
• Assessment of sensor performance in live-cell imaging.
• Benchmarking against traditional scaffold-bound fluorescent reporters.

Main Results:

• Genetically-encoded LLPS-sensors circumvent the need for direct IDP tagging.
• Tunable ultraweak scaffold-sensor interactions enable sensitive and innocuous probing.
• LLPS-sensors accurately report on condensate assembly and dynamics.

Conclusions:

• Epidermal LLPS-sensors offer a novel, high-fidelity approach for studying biomolecular condensates.
• These tools overcome limitations of traditional tagging methods.
• The developed sensors have broad applicability for intracellular condensate research across biological systems.