title: Directed evolution of a genetic circuit.
abstract: The construction of artificial networks of transcriptional control elements in living cells represents a new frontier for biological engineering. However, biological circuit engineers will have to confront their inability to predict the precise behavior of even the most simple synthetic networks, a serious shortcoming and challenge for the design and construction of more sophisticated genetic circuitry in the future. We propose a combined rational and evolutionary design strategy for constructing genetic regulatory circuits, an approach that allows the engineer to fine-tune the biochemical parameters of the networks experimentally in vivo. By applying directed evolution to genes comprising a simple genetic circuit, we demonstrate that a nonfunctional circuit containing improperly matched components can evolve rapidly into a functional one. In the process, we generated a library of genetic devices with a range of behaviors that can be used to construct more complex circuits.
authors.affiliation: Division of Chemistry and Chemical Engineering, California Institute of Technology, 210-41, 1200 East California Boulevard, Pasadena, CA 91125, USA.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Yokobayashi
authors.forename: Ron
authors.initials: R
authors.lastname: Weiss
authors.forename: Frances H
authors.initials: FH
authors.lastname: Arnold
mesh_terms: D004247:DNA / Q000378:metabolism; D019143:Evolution, Molecular; D005786:Gene Expression Regulation; D005818:Genetic Engineering; D016296:Mutagenesis; D010957:Plasmids; D012097:Repressor Proteins / Q000502:physiology
chemical_list: D012097:Repressor Proteins; D004247:DNA