Eukaryotic-like Synthetic Cells with Chemically Controlled Protein Localization

Area of Science:

• Synthetic Biology
• Cell Biology
• Biochemistry

Background:

• Organelle compartmentalization and protein localization are crucial for eukaryotic cell function.
• Developing bottom-up synthetic cell models is essential for understanding cellular processes.

Purpose of the Study:

• To create a bottom-up synthetic cell platform with chemically controlled protein localization within an artificial organelle.
• To demonstrate multistep cascade reactions and phenotype control in synthetic cells.

Main Methods:

• Construction of cell-sized liposomes (giant unilamellar vesicles, GUVs) encapsulating a DNA-droplet-based organelle.
• Utilizing proteins fused to Escherichia coli dihydrofolate reductase for inducible recruitment to the organelle interior.
• Employing a synthetic trimethoprim derivative for chemical control of protein localization.
• Coupling protein relocalization with a sequence-specific protease for cascade reactions.

Main Results:

• Achieved rapid, on-demand protein recruitment to the artificial organelle upon addition of a small molecule inducer.
• Demonstrated chemically induced, multistep cascade reactions including protein relocalization and organelle-specific enzymatic activity.
• Successfully controlled synthetic cell phenotypes, such as pore formation in the GUV membrane, through programmed reactions.

Conclusions:

• The developed system provides a versatile platform for creating eukaryotic-like synthetic cells.
• This approach enables sophisticated and programmable functions through controlled compartmentalization and reaction cascades.
• Offers a novel strategy for bottom-up construction of functional synthetic cells.