Enhanced Multiplex Genome Engineering Through Co-operative Oligonucleotide Co-selection

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2012 May 29

PMID 22638574

Citations 47

Authors

Peter A Carr

Harris H Wang

Bram Sterling

Farren J Isaacs

Marc J Lajoie

George Xu

George M Church

Joseph M Jacobson

Affiliations

Soon will be listed here.

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.

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