Long-lived protein expression in hydrogel particles: towards artificial cells

Area of Science:

• Synthetic Biology
• Biotechnology
• Materials Science

Background:

• Cell-free systems offer advantages for protein synthesis but lack long-term stability.
• Developing artificial systems that mimic cellular functions is crucial for advanced biotechnological applications.

Purpose of the Study:

• To create a novel cell-mimic particle system for sustained protein expression.
• To demonstrate the ability to regulate gene expression within these particles.
• To engineer an in vitro genetic oscillator using these particles.

Main Methods:

• Immobilization of cell-free transcription-translation factors onto hydrogel particle backbones.
• Encapsulation of plasmid DNA and ribosomes within hydrogel particles.
• Utilizing the lac operon system for inducible gene expression control.

Main Results:

• Achieved stable protein expression for at least 11 days with continuous nutrient supply.
• Demonstrated precise regulation of mCherry expression in response to isopropyl β-d-1-thiogalactopyranoside (IPTG) levels.
• Successfully constructed an in vitro genetic oscillator exhibiting protein feedback loops and oscillations.

Conclusions:

• The developed cell-mimic particles provide a robust platform for long-lived protein expression.
• These particles enable dynamic control over gene expression and the creation of synthetic gene circuits.
• Potential applications include gene circuit engineering, metabolic engineering, and biosensor development.