Tardigrade-Derived Strategy for Low-Cost Storage of Cell-Free Expression Lysates

Area of Science:

• Synthetic Biology
• Biochemistry
• Biotechnology

Background:

• Cell-free expression systems (CFESs) offer rapid prototyping but face limitations due to costly preparation and strict cold storage needs.
• Existing methods for stabilizing CFESs via lyophilization often compromise activity due to protective additives.

Purpose of the Study:

• To develop a stable, room-temperature desiccated CFES that overcomes current limitations.
• To investigate the protective role of tardigrade-derived cytosolic-abundant heat-soluble (CAHS) proteins in preserving CFES activity after drying.

Main Methods:

• Incorporation of tardigrade-derived CAHS protein into CFES lysates.
• Lyophilization of CFES with CAHS protein at room temperature.
• Assessment of protein synthesis activity, mRNA synthesis kinetics, and translation yields post-desiccation.

Main Results:

• CFES containing CAHS protein maintained protein synthesis activity after room-temperature desiccation without additional protective agents.
• Unprotected lysates showed impaired mRNA synthesis kinetics and reduced translation yields following desiccation.
• CAHS protein effectively protected the biosynthetic machinery during drying.

Conclusions:

• Tardigrade-derived CAHS protein enables the creation of stable, solid-state CFESs through low-cost room-temperature desiccation.
• This approach significantly enhances the accessibility and portability of CFESs for synthetic biology applications.
• Tardigrade proteins represent a promising resource for developing robust and user-friendly cell-free systems.