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Published on: January 11, 2011
Genome-scale promoter engineering by coselection MAGE
Harris H Wang1, Hwangbeom Kim, Le Cong
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. harris.wang@wyss.harvard.edu
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.
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