Year: 2025
Type: preprint
Abstract: Lipid membrane-bound synthetic cells provide programmable, cell-like compartments for on-demand biomolecule production. However, most cell-free gene expression systems in synthetic cells are regulated by user-imposed cues rather than signals associated with environmental or physiological states. Here, we present an acidity-transducing synthetic cell that converts external pH changes into nucleic acid information to drive in situ protein synthesis. The system integrates gramicidin A proton channels for pH sensing, triplex-forming single-stranded DNA (ssDNA) that releases a trigger ssDNA upon acidification, and a toehold switch RNA that activates translation in response to the released trigger ssDNA. This work further reveals that tuning the annealing length between the pH-responsive and trigger ssDNAs controls the trigger-release pH, which is critical for enabling acid-triggered protein synthesis. The synthetic cells retain their pH-responsive activity when embedded in alginate hydrogels, creating acidity-responsive materials that synthesize proteins in situ rather than release preloaded cargo. Cell-penetrating peptide tagging further enables selective protein release and target-specific binding. This work establishes a molecular transduction strategy for programming synthetic cell materials to sense environmentally relevant acidity and generate functional biomolecules on demand. Table of Contents A synthetic cell is engineered to convert environmental acidification into nucleic acid signaling that activates protein synthesis. By coupling proton-channel sensing, pH-responsive DNA transduction, and toehold-regulated translation, the platform enables acid-triggered protein production within hydrogel-embedded synthetic cells. This strategy expands the repertoire of condition-associated cues that synthetic cells can interpret for responsive biomaterials, biosensing, and controlled delivery.
Institutions: University of Michigan, Boston University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Awards: Development of a mechanosensitive synthetic cell for mediating intercellular communication., Shear stress-activated synthetic cells for targeted drug release in stenotic blood vessels, Model-guided design of RNA stabilizing elements for improved coronavirus diagnostics, G5520316217, Programming cellular behavior by mechanical forces, G9000167822, G9000183231, G9000189331, G9000189389, Sensing and modulating the chemokine environment with synthetic cells, Ideas Lab: CFIRE: Meta-PURE: End-Use-Driven Cell-Free Modules
OpenAlex ID: W4416407608