Principles of cell-free genetic circuit assembly.
Cell-free genetic circuit elements were constructed in a transcription-translation extract. We engineered transcriptional activation and repression cascades, in which the protein product of each stage is the input required to drive or block the following stage. Although we can find regions of linear response for single stages, cascading to subsequent stages requires working in nonlinear regimes. Substantial time delays and dramatic decreases in output production are incurred with each additional stage because of a bottleneck at the translation machinery. Faster turnover of RNA message can relieve competition between genes and stabilize output against variations in input and parameters.
Noireaux
Vincent
V
Center for Studies in Physics and Biology, The Rockefeller University, New York, NY 10021, USA. noireav@mail.rockefeller.edu
Bar-Ziv
Roy
R
Libchaber
Albert
A
D000818:Animals; D002474:Cell-Free System / Q000502:physiology*; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D005819:Genetic Markers; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins; D007700:Kinetics; D008156:Luciferases / Q000235:genetics; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D014158:Transcription, Genetic
D005819:Genetic Markers; D008164:Luminescent Proteins; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D008156:Luciferases