Ultrasonic Control of Protein Splicing by Split Inteins

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Sonopharmacology and sonogenetics utilize ultrasound to modulate biological processes.
• Controlling protein activity remotely is crucial for therapeutic interventions.

Purpose of the Study:

• To develop an ultrasound-responsive protein splicing system for spatiotemporal control of protein ligation.
• To demonstrate the system's ability to control protein synthesis and cellular signaling.

Main Methods:

• Engineered split inteins were caged and activated by ultrasound-triggered thrombin release.
• A DNA-based carrier system was used to deliver the caged components.
• The system's efficacy was tested using superfolder green fluorescence protein (GFP) and calcitonin synthesis in vitro.

Main Results:

• Achieved spatiotemporal control of split-intein-mediated protein ligation using focused ultrasound.
• Successfully synthesized GFP and calcitonin in a controlled manner.
• Triggered calcitonin receptor-mediated signal transduction in cells without affecting cell viability.

Conclusions:

• The developed system provides a general method for ultrasound-controlled protein function.
• This expands the sonogenetic toolbox with protein splicing technologies.
• Offers a potential pathway for remote control of protein functions in deep tissues.