https://pages.htgaa.org/2026a/srinidhi-vasan/homework/week-9-cell-free-systems/index.htmlExplain the main advantages of cell-free protein synthesis over traditional in vivo methods, specifically in terms of flexibility and control over experimental variables. Name at least two cases where cell-free expression is more beneficial than cell production. Flexibility and Control: Since there is no cell membrane, the system is an open environment, where we can directly manipulate concentrations of substrates, add non-canonical amino acids, or introduce specific inhibitors/activators with no worries about cellular transport or toxicity. Case 1: Toxic Proteins: Many proteins, such as antimicrobial peptides or certain enzymes, are lethal to host cells. CFPS allows the production of these proteins. Case 2: Rapid Prototyping: In vivo methods need time-consuming cloning, transformation, and cell culture. CFPS use’s linear DNA as a template, reducing the time from days to hours. Describe the main components of a cell-free expression system and explain the role of each component. Cell Extract (Crude Lysate): It contains ribosomes, aminoacyl-tRNA synthetases, translation factors, and tRNAs. Energy and Buffer Systems: This includes ATP and GTP , an energy regeneration substrate (like Phosphoenolpyruvate), and essential ions (Mg2+ and K+) to maintain enzymatic activity and pH. Genetic Template and Building Blocks: DNA provides the instructions, while the 20 standard amino acids provide the raw material for the protein chain. Why is energy provision regeneration critical in cell-free systems? Describe a method you could use to ensure continuous ATP supply in your cell-free experiment. Protein synthesis is energetically expensive. ATP is consumed rapidly for amino acid activation and ribosome movement, and it is also naturally degraded by other enzymes in the extract.Hugoen