pH-responsive synthetic cells for controlled protein synthesis and release

Area of Science:

• Synthetic biology
• Biochemistry
• Biomaterials

Background:

• Acidic pH is a key indicator of various pathological conditions, including cancer and infections.
• Developing responsive systems for biological applications is crucial for diagnostics and therapeutics.

Purpose of the Study:

• To engineer a synthetic cell with pH-responsive protein synthesis and release capabilities.
• To explore the integration of molecular modules for acid-triggered biological functions.
• To demonstrate the potential of pH-responsive synthetic cells in biomaterials and targeted protein delivery.

Main Methods:

• Integration of three molecular modules: a proton channel for pH sensing, pH-responsive single-stranded DNA (ssDNA) for trigger release, and a toehold switch RNA for translation activation.
• Optimization of annealing length between pH-responsive and trigger DNA strands for efficient acid-triggered protein synthesis.
• Embedding synthetic cells in hydrogels and coupling protein translation with cell-penetrating peptide technology.

Main Results:

• Successful demonstration of acid-responsive protein expression within synthetic cells.
• Identification of critical parameters (annealing length) for optimizing acid-triggered protein synthesis.
• Development of pH-responsive hydrogels and a system for selective protein release using cell-penetrating peptides.

Conclusions:

• Engineered synthetic cells can exhibit controlled protein synthesis and release in response to acidic pH.
• The developed system offers a versatile platform for creating smart biomaterials and targeted drug delivery systems.
• This work provides a foundation for developing advanced diagnostics and therapeutics for pH-associated diseases.