Cell-Free Protein Expression in Polymer Materials

Abstract

While synthetic biology has advanced complex capabilities such as sensing and molecular synthesis in aqueous solutions, important applications may also be pursued for biological systems in solid materials. Harsh processing conditions used to produce many synthetic materials such as plastics make the incorporation of biological functionality challenging. One technology that shows promise in circumventing these issues is cell-free protein synthesis (CFPS), where core cellular functionality is reconstituted outside the cell. CFPS enables genetic functions to be implemented without the complications of membrane transport or concerns over the cellular viability or release of genetically modified organisms. Here, we demonstrate that dried CFPS reactions have remarkable tolerance to heat and organic solvent exposure during the casting processes for polymer materials. We demonstrate the utility of this observation by creating plastics that have spatially patterned genetic functionality, produce antimicrobials in situ, and perform sensing reactions. The resulting materials unlock the potential to deliver DNA-programmable biofunctionality in a ubiquitous class of synthetic materials.

Cell-Free Gene Expression in Bioprinted Fluidic Networks

“…Such systems are expected to have advanced (bio)chemical sensing capabilities and the ability to programmably produce (bio)molecules on demand. A promising approach for the execution of such biochemical functions is to utilize cell-free technology, which has the advantage of not being restricted by the complex requirements of living systems and can be integrated in synthetic materials . Cell-free gene expression reactions are typically performed in cell extracts that are supplemented with biosynthetic building blocks (NTPs and amino acids) and biochemical cofactors (e.g., ATP, coenzyme A), allowing for high yield in situ production of proteins. − …”