Journal
NUCLEIC ACIDS RESEARCH
Volume 40, Issue 17, Pages -Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gks455
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Funding
- National Science Foundation (SynBERC) [0540879]
- Center for Bits and Atoms [0122419]
- Genes and Genomes Systems Cluster [0719344]
- Department of Energy (Genome to Life Center) [DE-FG02-03ER6344]
- Technology Development Fellowship from the Wyss Institute for Biologically Inspired Engineering
- National Institutes of Health [1DP5OD009172-01]
- US DOD NDSEG fellowship
- Massachusetts Institute of Technology, Cambridge, MA, USA
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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.
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