Abstract
Cell-free protein synthesis (CFPS) system is a simple, rapid, and sensitive tool that is devoid of membrane-bound barriers, yet contains all the mandatory substrates, biomolecules, and machineries required for the synthesis of the desired proteins. It has the potential to overcome loopholes in the current in vivo production systems and is a promising tool in both basic and applied scientific research. It facilitates a simplified organization of desired experiments with a variety of reaction conditions, making CFPS a powerful tool in biological research. It has been used for the expansion of genetic code, assembly of viruses, and in metabolic engineering for production of toxic and complex proteins. Subsequently, CFPS systems have emerged as potent technology for high-throughput production of membrane proteins, enzymes, and therapeutics. The present review highlights the recent advances and uses of CFPS systems in biomedical, therapeutic, and biotechnological applications. Additionally, we highlight possible solutions to the potential biosafety issues that may be encountered while using CFPS technology.
Highlights
Synthetic biology has emerged and continues to grow as a burgeoning scientific field that combines engineering principles with the biological sciences
Synthetic biology is a modern and innovative scientific discipline with an aim to improve the existing industrial practices, addressing issues of poor yields and poor cost-to-product ratios, as well as the problems of current practices that inevitably damage our ecosystems through polluting acts
In any of these cases, it would be prudent to consider alternatives, and it is in synthetic biology that novel and alternative routes for the fabrication of many value-added products have been found with a compelling amount of accomplishments and an ever-fertile basis to grow future products and industries
Summary
Synthetic biology has emerged and continues to grow as a burgeoning scientific field that combines engineering principles with the biological sciences. The daunting complexity and barrier rendered by the cell membrane prompt numerous difficulties, such as experiment being hard to standardize, incompatibility issues, and variability To address these issues, cell-free protein synthesis (CFPS) systems, known as in vitro protein synthesis, have emerged as a key tool that can work without the use of living cells. CFPS represents a historically important component in the field of biochemistry, duly acknowledging the pioneering effort made by Nobel laureate Eduard Buchner (Nobel Prize in Chemistry 1907) for the discovery of fermentation in yeast cell extracts (YCE) (Buchner, 1897) It has since been repurposed for the understanding of biological processes, most notably contributing to the discovery of genetic code through the use of Escherichia coli cell extract by Nirenberg and colleagues (Nirenberg and Matthaei, 1961; Matthaei et al, 1962), which led them to win and share the Nobel Prize for Physiology or Medicine in year 1968, together with Har Gobind Khorana and Robert Holley. We highlight the recent progress and uses of CFPS in biomedical, therapeutic, industrial, and biotechnological applications
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