Imperial/Cell-Free/Comparison

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Cell-Free: A Comparison of In Vivo and In Vitro Systems

In-Vitro Expression Systems
In-Vivo Expression Systems
Non-infectious because of non-proliferative nature Some strains may be pathogenic
Process is quick and simple requiring only preparation of cell extract and feeding solution and subsequent addition of DNA template Process is laborious involving DNA cloning and transformation and protein expression
Good control can be achieved easily using modified reaction conditions such as addition of accessory elements or inhibitory factors Less controllability because of the presence of endogenous substances and because cells do not survive extreme conditions
Both plasmid and linear DNAs and can be used as templates for expression Plasmid DNAs are usually used. Linear DNAs are easily degraded by endogenous nucleases
Protein degradation is minimized by adding protease inhibitors Synthesized proteins may be degraded by endogenous proteases
Toxic proteins can be synthesized in large quantities Synthesis of toxic proteins may kill the cells
Proteins containing unnatural amino acids can be achieved Difficult to produce proteins containing unnatural amino acids
Shorter lifespan since system cannot replicate Longer lifespan since system can replicate
More expensive because of the constant need for nutrient and energy supply Less expensive because of the ability of the system to generate energy from relatively cheap nutrient source
Less characterized, less experience of use in the laboratories Better characterized, more experience of use in the laboratories



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