A PEGDA/DNA Hybrid Hydrogel for Cell-Free Protein Synthesis

Frontiers in Chemistry
Jinhui CuiDan Luo

Abstract

Cell-free protein synthesis (CFPS) has the advantage of rapid expression of proteins and has been widely implemented in synthetic biology and protein engineering. However, the critical problem limiting CFPS industrial application is its relatively high cost, which partly attributes to the overexpense of single-use DNA templates. Hydrogels provide a possible solution because they can preserve and reutilize the DNA templates in CFPS and have great potential in elevating the protein production yield of the CFPS. Here, we presented a low-cost hybrid hydrogel simply prepared with polyethylene glycol diacrylate (PEGDA) and DNA, which is capable of high-efficient and repeated protein synthesis in CFPS. Parameters governing protein production specific to hybrid hydrogels were optimized. Structures and physical properties of the hybrid hydrogel were characterized. Transcription and expression kinetics of solution phase system and gel phased systems were investigated. The results showed that PEGDA/DNA hydrogel can enhance the protein expression of the CFPS system and enable a repeated protein production for tens of times. This PEGDA/DNA hybrid hydrogel can serve as a recyclable gene carrier for either batch or continuous protein expressi...Continue Reading

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Citations

Jul 9, 2020·Synthetic and Systems Biotechnology·Marilyn S LeeMatthew W Lux
Apr 13, 2021·ACS Synthetic Biology·Steven M BlumMatthew W Lux
Jun 22, 2021·Frontiers in Bioengineering and Biotechnology·Xueting JianYang Liu
Aug 11, 2021·Polymers·Alessa SchippergesChristof M Niemeyer
Nov 11, 2020·ACS Synthetic Biology·Ana I Benítez-MateosFernando López-Gallego
Jan 16, 2021·ACS Biomaterials Science & Engineering·Vinod MoryaDhiraj Bhatia

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Methods Mentioned

BETA
PCR
scanning electron microscopy
rheology
two hybrid

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