Constructing Cell-Free Expression Systems for Low-Cost Access.

Area of Science:

• Biotechnology
• Synthetic Biology
• Molecular Biology

Background:

• Cell-free systems (CFS) are valuable for studying genetic circuits and teaching, but high costs limit accessibility.
• Expensive reagents and specialized equipment hinder widespread adoption, particularly in low- and middle-income countries.

Purpose of the Study:

• To develop low-cost, accessible strategies for preparing and distributing cell-free gene expression systems.
• To overcome economic and logistical barriers preventing global use of CFS technology.

Main Methods:

• Explored low-cost silica bead-based drying for cell-free reaction stability.
• Investigated lactose as a protein synthesis enhancer in maltodextrin-based CFS.
• Demonstrated endogenous nucleotide triphosphate regeneration in CFS using PEP and maltodextrin.

Main Results:

• Developed a low-cost method for producing stable, desiccated cell-free extracts.
• Identified lactose as an effective additive for enhancing protein synthesis.
• Achieved high yields of sfGFP expression using a cell-free system free of exogenous nucleotide triphosphates.
• Produced active high-value reagents like *Bst* polymerase and *Bsa*I restriction enzyme.

Conclusions:

• The developed modifications result in desiccated cell-free extracts that are 203-424 times cheaper than commercial versions.
• These advancements significantly improve the accessibility and affordability of cell-free systems for research and education worldwide.