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Raig, R. M.

Publications and source records attributed to Raig, R. M..

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Cell-Free Protein Expression in Polymer Materials

While synthetic biology has advanced complex capabilities like sensing and molecule 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 makes 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 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 bio-functionality in a ubiquitous class of synthetic materials.

synthetic biology↗

Lyophilized cell-free systems display tolerance to organic solvent exposure

Cell-free systems offer a powerful way to deliver biochemical activity to the field without cold chain storage. These systems are capable of sensing as well as biosynthesis of useful molecules at the point of need. So far, cell-free protein synthesis (CFPS) reactions have been studied as aqueous solutions in test tubes or absorbed into paper or cloth. Embedding biological functionality into broadly-used materials, such as plastic polymers, represents an attractive goal. Unfortunately, this goal has for the most part remained out of reach, presumably due to the fragility of biological systems outside of aqueous environments. Here, we describe a surprising and useful feature of lyophilized cell-free lysate systems: tolerance to a variety of organic solvents. Screens of individual CFPS reagents and different CFPS methods reveal that solvent tolerance varies by CFPS reagent composition. Tolerance to suspension in organic solvents may facilitate the use of polymers to deliver dry cell-free reactions in the form of coatings or fibers, or allow dosing of analytes or substrates dissolved in non-aqueous solvents, among other processing possibilities.Competing Interest StatementThe authors have declared no competing interest.AbbreviationsCFPScell-free protein synthesisGFPgreen fluorescent proteinDCMdichloromethaneDMFdimethylformamideDMSOdimethylsulfoxideTHFtetrahydrofuranView Full Text

synthetic biology↗