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Enhanced multiplex genome engineering through co-operative oligonucleotide co-selection

Author(s)
Carr, Peter A., Sr.; Wang, Harris H.; Sterling, Bram Henry; Isaacs, Farren J.; Lajoie, Marc J.; Xu, George Jing; Church, George M.; Jacobson, Joseph; ... Show more Show less
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Abstract
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.
Date issued
2012-05
URI
http://hdl.handle.net/1721.1/71773
Department
Harvard University--MIT Division of Health Sciences and Technology; Massachusetts Institute of Technology. Center for Bits and Atoms; Program in Media Arts and Sciences (Massachusetts Institute of Technology)
Journal
Nucleic Acids Research
Publisher
Oxford University Press (OUP)
Citation
Carr, P. A. et al. “Enhanced Multiplex Genome Engineering Through Co-operative Oligonucleotide Co-selection.” Nucleic Acids Research (2012).
Version: Final published version
ISSN
0305-1048
1362-4962

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