Engineered PROTAC-CID Systems for Mammalian Inducible Gene Regulation

Area of Science:

• Molecular Biology
• Synthetic Biology
• Gene Therapy

Background:

• Chemically induced dimerization (CID) is crucial for biomedical research but faces limitations in scope and in vivo application.
• Existing CID tools lack versatility and scalability for complex gene regulation tasks.

Purpose of the Study:

• To engineer proteolysis-targeting chimera-based scalable CID (PROTAC-CID) platforms for inducible gene regulation and editing.
• To expand the utility of CID tools in human cells and in vivo models.

Main Methods:

• Systematic engineering of PROTAC systems to create PROTAC-CID platforms.
• Development of orthogonal PROTAC-CIDs for gradient gene expression control and multiplexing.
• Integration of PROTAC-CID with genetic circuits for inducible DNA editor expression.
• Packaging PROTAC-CID systems into adeno-associated viral vectors for in vivo applications.

Main Results:

• Demonstrated scalable PROTAC-CID platforms for inducible gene regulation and editing.
• Achieved fine-tuned gene expression and multiplexed biological signals using orthogonal PROTAC-CIDs.
• Enabled transient genome manipulation via digitally inducible DNA recombinases and editors.
• Showcased inducible and reversible gene activation in vivo using AAV-packaged PROTAC-CID systems.

Conclusions:

• The PROTAC-CID platform offers a versatile molecular toolbox for advanced gene regulation.
• This technology significantly expands the possibilities of chemically inducible gene control in mammalian systems.
• The developed tools hold promise for future therapeutic applications in gene therapy and beyond.