To advance the understanding and development of LLPS-sensitive tools, we pursued new quantitative approaches to fully dissect the biomolecular engineering and properties of epidermal LLPS-sensors. Excitingly, our new experiments and data demonstrated two notable properties of LLPS-sensors: (i) highly tunable range of sensitivity, and (ii) innocuous probing of concentration-dependent intracellular LLPS dynamics. Benchmarking our top-performing epidermal LLPS-sensor against a scaffold-bound fluorescent reporter-client, we discovered that ultraweak scaffold-sensor interactions are key to the high-fidelity probing of nascent and established biomolecular condensates. Together with a detailed biomolecular framework to engineer scaffold-specific LLPS-sensors, our new findings demonstrate a path toward rigorous intracellular probing of IDP-governed biomolecular condensates across biological systems.
The paper just appeared online in ACS Sensors. The work was led by Alexa Avecilla, with outstanding contributions from Jeremy Thomas.