Programmable synthetic cell networks regulated by tuneable reaction rates

Area of Science:

• Synthetic Biology
• Chemical Kinetics
• Biophysics

Background:

• Molecular diffusion and compartmentalisation are crucial for biological information processing and network dynamics.
• Controlling these processes is key for designing and managing reaction networks.
• Understanding compartmentalisation effects on DNA reactions is essential for synthetic biology applications.

Purpose of the Study:

• To investigate the impact of compartmentalisation on autocatalytic PEN DNA reactions within proteinosomes.
• To characterise unique reaction behaviours not observed in bulk conditions.
• To exploit these behaviours for regulating reaction kinetics in synthetic biological networks.

Main Methods:

• Integration of PEN DNA reactions into semi-permeable proteinosomes.
• Characterisation of reaction kinetics under compartmentalised versus bulk conditions.
• Bottom-up construction of two-node reaction networks using linear and autocatalytic reactions.

Main Results:

• Compartmentalisation within proteinosomes significantly increased PEN DNA reaction rates (by an order of magnitude).
• Reaction kinetics in proteinosomes were more readily tunable by enzyme concentrations compared to buffer solutions.
• Unique behaviours, not seen in bulk, were observed in compartmentalised systems.

Conclusions:

• Semi-permeable proteinosomes provide a platform for enhanced and tunable PEN DNA reactions.
• Compartmentalisation offers a powerful strategy for controlling reaction kinetics in synthetic biological systems.
• This work advances the bottom-up design of complex, network-based biological functions.