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

Enhanced multiplex genome engineering through co-operative oligonucleotide co-selection.
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DOI:
10.1093/nar/gks455
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发表时间:
2012-09-01
影响因子:
14.9
通讯作者:
Jacobson JM
Jacobson JM
中科院分区:
生物学2区
文献类型:
--
作者:
Carr PA;Wang HH;Sterling B;Isaacs FJ;Lajoie MJ;Xu G;Church GM;Jacobson JM

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生物体的基因组规模工程需要精确和经济的方法来有效地修改染色体内的许多位点。其中一个例子是在复制过程中将化学合成的单链脱氧核糖核酸(寡核苷酸)定向整合到大肠杆菌的染色体中。在此,我们提出了一种在多重基因组工程中产生高度修饰细胞的一般共选择策略。我们证明,整个基因组的不同位点可以通过利用目标位点500 kb内的可选择标记轻松地同时进行修饰。我们将该技术应用于大肠杆菌基因组的80个位点的修饰。
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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