title: Rationally designed bidirectional promoter improves the evolutionary stability of synthetic genetic circuits.
abstract: One problem with synthetic genes in genetically engineered organisms is that these foreign DNAs will eventually lose their functions over evolutionary time in absence of selective pressures. This general limitation can restrain the long-term study and industrial application of synthetic genetic circuits. Previous studies have shown that because of their crucial regulatory functions, prokaryotic promoters in synthetic genetic circuits are especially vulnerable to mutations. In this study, we rationally designed robust bidirectional promoters (BDPs), which are self-protected through the complementarity of their overlapping forward and backward promoter sequences on DNA duplex. When the transcription of a target non-essential gene (e.g. green fluorescent protein) was coupled to the transcription of an essential gene (e.g. antibiotic resistance gene) through the BDP, the evolutionary half-time of the gene of interest increases 4-10 times, depending on the strain and experimental conditions used. This design of using BDPs to increase the mutational stability of genetic circuits can be potentially applied to synthetic biology applications in general.
authors.affiliation: Genomics Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Mail Stop 84-363, Berkeley, CA 94720, USA. songyang@lbl.gov
authors.forename: Song
authors.initials: S
authors.lastname: Yang
authors.forename: Sean C
authors.initials: SC
authors.lastname: Sleight
authors.forename: Herbert M
authors.initials: HM
authors.lastname: Sauro
mesh_terms: D004926:Escherichia coli / Q000235:genetics; D019143:Evolution, Molecular; D005786:Gene Expression Regulation; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics; D007613:Kanamycin Resistance / Q000235:genetics; D009154:Mutation; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
chemical_list: D049452:Green Fluorescent Proteins