- Harshini Davuluri
Also known as in-vitro protein synthesis, CFPS uses biological machinery in a cell-free system. It is a sensitive and straightforward tool that does not contain membrane-bound barriers but has all the required components such as an energy source, cell extracts, supply of amino acids, cofactors such as magnesium, and desired DNA (plasmids or linear expression templates). The energy source is an essential part of a cell-free reaction. Generally, phosphoenolpyruvate, creatine phosphate, and acetyl phosphate are the sources. Since a homeostatic condition does not constrain this environment, it can control the translation environment, co-solubilization of membrane proteins, and selective and site-specific labelling.
CFPS represents a historically important component within biochemistry, duly acknowledging the pioneering effort made by Nobelist Buchner (Nobel Prize in Chemistry 1907) for the invention of fermentation in yeast cell extracts. Through the utilization of E. coli extract, Nirenberg and colleagues had developed an understanding, for which they received a Nobel Prize in 1968. These days, the primary sources of cell extracts are from E. coli (ECE), wheat germ (WGE), insect cell (ICE). Being an inexpensive extract, ECE is the most popular lysate that takes the least time to create and can provide high protein yields.
Cell-free protein synthesis is now emerging as a powerful technology for the high-throughput production of membrane proteins, enzymes, and therapeutics. Since live cells are not used, there is a negligible level of toxicity. This advantage enables numerous applications such as inserting modified tRNA and unnatural amino acids required for such a reaction.
Recent research in the University of Warwick, United Kingdom, has shown that the advances in such areas of synthetic biology have extended their use in basic sciences, biomedical sciences, biotechnology, and industries. This expansion and development have become particularly useful for accelerating the invention and innovation in synthetic biology.
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