title: Genome-scale promoter engineering by coselection MAGE.
abstract: Multiplex automated genome engineering (MAGE) uses short oligonucleotides to scarlessly modify genomes; however, insertions >10 bases are still inefficient but can be improved substantially by selection of highly modified chromosomes. Here we describe 'coselection' MAGE (CoS-MAGE) to optimize biosynthesis of aromatic amino acid derivatives by combinatorially inserting multiple T7 promoters simultaneously into 12 genomic operons. Promoter libraries can be quickly generated to study gain-of-function epistatic interactions in gene networks.
authors.affiliation: Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. harris.wang@wyss.harvard.edu
authors.forename: Harris H
authors.initials: HH
authors.lastname: Wang
authors.forename: Hwangbeom
authors.initials: H
authors.lastname: Kim
authors.forename: Le
authors.initials: L
authors.lastname: Cong
authors.forename: Jaehwan
authors.initials: J
authors.lastname: Jeong
authors.forename: Duhee
authors.initials: D
authors.lastname: Bang
authors.forename: George M
authors.initials: GM
authors.lastname: Church
mesh_terms: D057205:Automation, Laboratory; D017123:Bacteriophage T7 / Q000235:genetics; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D053263:Gene Regulatory Networks; D005818:Genetic Engineering / Q000379:methods*; D016678:Genome; D023281:Genomics; D007203:Indigo Carmine; D007211:Indoles / Q000378:metabolism; D009838:Oligodeoxyribonucleotides; D011401:Promoter Regions, Genetic / Q000235:genetics*
chemical_list: D007211:Indoles; D009838:Oligodeoxyribonucleotides; D007203:Indigo Carmine