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Can toxic proteins be produced easily? Explore the superpowers of CFPS!
In the biology community, cell-free protein synthesis (Cell-Free Protein Synthesis, referred to as CFPS) is attracting increasing attention. This technology allows scientists to produce proteins without living cells, and its potential is being fully explored. Today, we will explore in-depth the working principles, advantages, applications and possibilities of CFPS.
Basic Principles of CFPS
Cell-free protein synthesis is a process of synthesizing proteins in a controlled environment using the biomechanism of cells. Compared with traditional live cell protein synthesis, CFPS is not limited by cell membranes and survival status.
CFPS combines various biochemical components such as cell extracts, energy sources, amino acid supplies, and DNA carrying the desired genes.
These components are taken from target cells, and the necessary cellular mechanisms are obtained through the destruction of the cell structure and centrifugation process, including ribosomes, aminoacyl-tRNA synthetase and other translation-related factors.
Advantages of CFPS
The advantages of CFPS are obvious, first of all, time efficiency. The process of preparing cell extracts usually takes only 1 to 2 days, while protein expression in living cells may take 1 to 2 weeks. In addition, the open reaction environment of CFPS enables direct manipulation of chemical conditions, allowing easy adjustment of concentration and sampling.
Another advantage of CFPS is the ability to produce toxic proteins, which is relatively difficult in living cells.
This is especially important because certain desired proteins can cause toxicity to cells during synthesis, and CFPS does not need to worry about this.
CFPS Application
CFPS has potential applications in many fields, including the synthesis of protein structures of unnatural amino acids, which is particularly critical for synthetic biology.
With CFPS, we can explore the boundaries of synthetic biology, including protein evolution, nanomachines, and virus-like particle synthesis for vaccines and drug therapies.
Limitations and Challenges
Although CFPS has many advantages, it also faces challenges, especially the problem of DNA degradation. In cell extracts, endogenous nucleases may attack rectangular expression templates (LETs), which makes LETs more vulnerable than plasmids.
The researchers are working to increase the yield of LETs by using nuclease-inhibiting proteins, such as the bacteriophage lambda gam protein, and make it similar to the yield of plasmids.
Types of cell-free systems
The commonly used cell-free extracts today come from different sources, such as E. coli, rabbit red blood cells, wheat germ, etc. Each extract has its own unique advantages and disadvantages, with E. coli extracts favored for their economic and high yields.
History of CFPS
Cell-free protein synthesis technology has a history of more than 60 years. As early as 1961, Marshall Nirenberg and Heinrich J Matai performed the first coding assay at the National Institutes of Health and demonstrated the potential of CFPS for the first time. They used CFPS to translate a polyuracil RNA sequence and found that the synthetic polypeptide only contains phenylalanine, which infers that UUU encodes phenylalanine.
With the continuous advancement of technology, the technology and application of cell-free protein synthesis will become more extensive, and provide possible solutions to various biomedical and high-tech industries. However, as the field expands, we can’t help but wonder whether CFPS can create a new chapter in life sciences in the future and become a key to solving human health problems?
