Open AccessEnhanced multiplex genome engineering through co-operative oligonucleotide co-selection
Author(s) -
Peter A. Carr,
Harris H. Wang,
Bram Sterling,
Farren J. Isaacs,
Marc J. Lajoie,
George Xu,
George M. Church,
Joseph M. Jacobson
Publication year - 2012
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/gks455
Subject(s) - biology , genome , multiplex , oligonucleotide , genome engineering , computational biology , genetics , dna , selection (genetic algorithm) , escherichia coli , chromosome , gene , genome editing , computer science , artificial intelligence
Genome-scale engineering of living organisms requires precise and economical methods to efficiently modify many loci within chromosomes. One such example is the directed integration of chemically synthesized single-stranded deoxyribonucleic acid (oligonucleotides) into the chromosome of Escherichia coli during replication. Herein, we present a general co-selection strategy in multiplex genome engineering that yields highly modified cells. We demonstrate that disparate sites throughout the genome can be easily modified simultaneously by leveraging selectable markers within 500 kb of the target sites. We apply this technique to the modification of 80 sites in the E. coli genome.United States. Dept. of Energy. Genomes To Life (DE-FG02-03ER6344)National Science Foundation (U.S.). Genes and Genomes Systems Cluster (0719344)National Science Foundation (U.S.). Center for Bits and Atoms (0122419)National Science Foundation (U.S.). Synthetic Biology Engineering Research Center (0540879
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