A cell-free protein-producing gel

Abstract

Proteins are important biomaterials and are generally produced in living cells. Here, we show a novel DNA hydrogel that is capable of producing functional proteins without any living cells. This protein-producing gel (termed 'the P-gel system' or 'P-gel') consists of genes as part of the gel scaffolding. This is the first time that a hydrogel has been used to produce proteins. The efficiency was about 300 times higher than current, solution-based systems. In terms of volumetric yield, the P-gel produced up to 5 mg ml(-1) of functional proteins. The mechanisms behind the high efficiency and yield include improved gene stability, higher local concentration and a faster enzyme turnover rate due to a closer proximity of genes. We have tested a total of 16 different P-gels and have successfully produced all 16 proteins including membrane and toxic proteins, demonstrating that the P-gel system can serve as a general protein production technology.

Development of DNA Based Active Macro–Materials for Biology and Medicine: A Review

“…The potential of applying this hydrogel for drug delivery application has also been demonstrated. Very recently, this group also reported that by incorporating linear plasmids into polymer network, this hydrogel is capable of generating natural proteins under cell-free conditions (Park et al 2009). By using the similar Y-shaped DNA building blocks, but a different mechanism to connect these building blocks, Cheng group also put forth a hydrogel design based entirely on DNA nanostructures (Cheng et al 2009).…”

“…Additionally, DNA gels have been shown to be an ideal candidate for cell capsulation (Um et al 2006b), potentially, serving as in vivo protein factory for protein synthesis and delivery (Park et al 2009). Examples of the studies using DNA-only, DNA-as-backbone, and DNA crosslinked macromaterials on potential drug or gene delivery applications and the kinetics of release are in Figure 7.…”

Solid-Phase Cell-Free Protein Synthesis to Improve Protein Foldability

“…Later, the immobilization of biomolecule and synthetic solid-phase approaches have been successfully aided research for a widespread applications for both pre-translated biomolecules such as RNA/DNA and post-translated biomolecules such as protein/enzymes including SNP genotyping [16], DNA amplification [17], differential display [18], in vitro transcription [19], immunoassay [20,21], and others while promoted the development of microfabrication [22,23], high-throughput screening and automation strategies in many areas including proteomics. Very recently, a hydrogel-based system was introduced that improved the efficiency of CFPS up to 300 times than solution phase-based system [24].…”

Branched DNA-based Synthesis of Fluorescent Silver Nanocluster

“…Dendrimer-like DNAs such as X-shaped DNA (X-DNA) and Y-shaped DNA (Y-DNA) have attracted great interest for using DNAs as a generic instead of a genetic material. 24,25 These DNA building blocks can be readily incorporated into larger structures in a controlled manner and the controlled assembly is robust and efficient yielding stable and almost monodisperse entities. Herein, we report X-DNA and Y-DNA templated syntheses of Ag NCs and their optical properties.…”