Computational design of chemogenetic and optogenetic split proteins

Area of Science:

• Molecular Biology
• Synthetic Biology
• Biotechnology

Background:

• Chemogenetics and optogenetics are powerful tools for studying rapid biological processes by controlling protein activity.
• Split protein systems offer potential for controlling diverse protein functions but face challenges like spontaneous assembly and inefficient reassembly.
• Identifying optimal split sites and achieving efficient, inducible reassembly are critical hurdles in split protein design.

Purpose of the Study:

• To develop an automated approach for designing effective split proteins regulated by chemical ligands or light.
• To overcome limitations of existing split protein technologies, including spontaneous assembly and inefficient reassembly.
• To provide a user-friendly tool for researchers to design split proteins for various applications.

Main Methods:

• Developed a scoring function and an engineered domain to enhance protein half reassembly efficiency and minimize spontaneous assembly.
• Created an automated computational approach named SPELL (Split Proteins Enabled by Ligand or Light) for predicting split sites.
• Validated the SPELL approach by successfully designing and testing split proteins in living cells.

Main Results:

• Demonstrated high efficiency and reduced spontaneous assembly in split protein reassembly using the engineered domain and scoring function.
• Successfully applied the SPELL system to various proteins of different sizes and shapes in living cells.
• The SPELL server provides automated prediction of split sites, facilitating split protein design.

Conclusions:

• SPELL offers an automated and efficient method for designing split proteins for chemogenetic and optogenetic control.
• This technology has broad applicability for controlling diverse protein activities in biological research.
• The SPELL server democratizes the design of split proteins, accelerating research in synthetic biology and beyond.