Genome-scale promoter engineering by coselection MAGE

Nat Methods. 2012 Jun;9(6):591-3. doi: 10.1038/nmeth.1971. Epub 2012 Apr 8.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Automation, Laboratory
  • Bacteriophage T7 / genetics
  • Base Sequence
  • Chromosomes, Bacterial
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Regulatory Networks
  • Genetic Engineering / methods*
  • Genome
  • Genomics
  • Indigo Carmine
  • Indoles / metabolism
  • Oligodeoxyribonucleotides
  • Promoter Regions, Genetic / genetics*


  • Indoles
  • Oligodeoxyribonucleotides
  • Indigo Carmine