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

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.

SUBMITTER: Carr PA

PROVIDER: S-EPMC3458525 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature

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

Carr Peter A PA Wang Harris H HH Sterling Bram B Isaacs Farren J FJ Lajoie Marc J MJ Xu George G Church George M GM Jacobson Joseph M JM

Nucleic acids research 20120525 17

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 ...[more]

PMID: 22638574

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