https://pages.htgaa.org/2026a/giulia-sironi/homework/week-9-cell-free-systems/index.htmlcomplete controll of the system, no limit of cellular variability, expression is faster and you can add and remove stuff in real life. protein can be toxic for the cell and some are difficult to express in vivo cellukar extract, dna template, aminoacid, nucleotide to trascribe mrna , atp and gtp , buffer like mg k and ph to stabilize anzime and e ribosom and chaperoni or othe racessory stuff to have correct folding and energetic regeneration is critical bc ATP is crucial and widly use, example can be PEP or creatin + phosphate, or glycolycic like pathways. Prokaryotic vs eukaryotic CFPS Prokaryotic (E. coli extract) limitat folding but cheap and fast GFP o baterial enzymes (es. T7 RNA polymerase) Eukaryotic (rabbit reticulocyte / insect / mammalian extract) support complex folding and glycolysation slow and expensive IgG for exampleHugoen