Optogenetic control of protein binding using light-switchable nanobodies

Area of Science:

• Biotechnology
• Molecular Biology
• Optogenetics

Background:

• Optogenetic tools enable reversible control of engineered protein interactions.
• Limited tools exist for light-switchable binding to generic target proteins.
• A versatile tool for photoswitchable binding to diverse targets is needed.

Purpose of the Study:

• To develop a versatile class of chimeric photoswitchable proteins for controlling protein binding.
• To create opto-nanobodies (OptoNBs) with light-tunable binding capabilities.
• To demonstrate the utility of OptoNBs in various biological applications.

Main Methods:

• Engineering chimeric photoswitchable proteins (OptoNBs).
• Testing OptoNBs for light-dependent binding to target proteins.
• Evaluating OptoNBs in cellular and in vitro applications.

Main Results:

• OptoNBs demonstrate enhanced or inhibited binding to proteins of interest upon blue light illumination.
• OptoNBs successfully bind reversibly to endogenous intracellular targets.
• OptoNBs modulate signaling pathway activity and control binding to purified proteins in vitro.

Conclusions:

• Opto-nanobodies (OptoNBs) offer a versatile platform for programmable photoswitchable regulation of protein binding.
• OptoNBs facilitate applications such as controlling intracellular targets and protein purification.
• This technology advances the development of optogenetic tools for diverse protein targets.