title: Combinatorial engineering of intergenic regions in operons tunes expression of multiple genes.
abstract: Many applications of synthetic biology require the balanced expression of multiple genes. Although operons facilitate coordinated expression of multiple genes in prokaryotes and eukaryotes, coordinating the many post-transcriptional processes that determine the relative levels of gene expression in operons by a priori design remains a challenge. We describe a method for tuning the expression of multiple genes within operons by generating libraries of tunable intergenic regions (TIGRs), recombining various post-transcriptional control elements and screening for the desired relative expression levels. TIGRs can vary the relative expression of two reporter genes over a 100-fold range and balance expression of three genes in an operon that encodes a heterologous mevalonate biosynthetic pathway, resulting in a sevenfold increase in mevalonate production. This technology should be useful for optimizing the expression of multiple genes in synthetic operons, both in prokaryotes and eukaryotes.
authors.affiliation: Department of Chemical Engineering, University of California-Berkeley, Berkeley, CA 94720-1462, USA.
authors.forename: Brian F
authors.initials: BF
authors.lastname: Pfleger
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Pitera
authors.forename: Christina D
authors.initials: CD
authors.lastname: Smolke
authors.forename: Jay D
authors.initials: JD
authors.lastname: Keasling
mesh_terms: D000465:Algorithms; D021901:DNA, Intergenic / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics*; D015723:Gene Library; D018014:Gene Transfer Techniques; D005818:Genetic Engineering / Q000379:methods*; D009876:Operon / Q000235:genetics*
chemical_list: D021901:DNA, Intergenic