Synthetic protein degradation circuits using programmable cleavage and ligation by Sortase A

Area of Science:

• Synthetic biology
• Molecular biology
• Biotechnology

Background:

• BioPROTACs are heterobifunctional proteins for targeted protein degradation (TPD).
• TPD is valuable for probing protein functions and offers therapeutic potential for diseases.
• Current TPD methods primarily focus on protein attenuation.

Purpose of the Study:

• To introduce a synthetic framework for logic-gated, switchable TPD.
• To achieve conditional control of protein levels beyond simple degradation.
• To expand the applications of TPD in mammalian cells.

Main Methods:

• Developed a synthetic framework for logic-gated, switchable TPD.
• Exploited Sortase A (SrtA) functionalities for control input.
• Integrated SrtA input with protease gating for layered control.
• Utilized a Logic-gated AdPROM deploying SrtA-mediated Element Recombination (LASER) platform.

Main Results:

• Demonstrated switchable TPD using SrtA as a control input.
• Achieved conditional degradation phenotypes through protease gating.
• Expanded protein degradation outcomes using Boolean logic operations.
• Showcased flexibility in modulating native intracellular protein levels.

Conclusions:

• The LASER platform enables sophisticated, conditional control of protein levels in mammalian cells.
• This approach expands the utility of TPD for various applications.
• Potential applications include therapy, diagnostics, and biotechnology.