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Abstract
Harris H Wang1,2,6,*,Hwangbeom Kim3,6, Le Cong1,4,5, Jaehwan Jeong3, Duhee Bang3,* & George M Church1,4
1Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA. 2Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA. 3Department of Chemistry, Yonsei University, Seoul, Korea. 4Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. 5Program in Biological and Biomedical Sciences, Harvard Medical School, Boston, Massachusetts, USA. 6These authors contributed equally to this work.
*Correspondence to: Harris H Wang or Duhee Bang
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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