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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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.
DepartmentHarvard 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)
Nucleic Acids Research
Oxford University Press (OUP)
Carr, P. A. et al. “Enhanced Multiplex Genome Engineering Through Co-operative Oligonucleotide Co-selection.” Nucleic Acids Research (2012).
Final published version