Multigene Editing in the Escherichia coli Genome via the CRISPR-Cas9 System
Multigene Editing in the Escherichia coli Genome via the CRISPR-Cas9 System
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DOI：
10.1128/aem.04023-14
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发表时间：
2015-04-01
影响因子：
4.4
通讯作者：
Yang, Sheng
中科院分区：
文献类型：
--
作者：
Jiang, Yu;Chen, Biao;Yang, Sheng
构建工业上有用的微生物需要一种有效的基因组规模编辑工具。我们描述了一种靶向的、连续的多基因编辑策略，该策略通过使用化脓性链球菌II型CRISPR-Cas9系统应用于大肠杆菌基因组，以实现各种精确的基因组修饰，包括基因缺失和插入，最高效率为100%，能够实现多达三个靶标的同时多基因编辑。该系统还证明了成功的靶向染色体缺失柠檬酸塔图姆氏菌，另一种肠杆菌科，效率最高的100%。
An efficient genome-scale editing tool is required for construction of industrially useful microbes. We describe a targeted, continual multigene editing strategy that was applied to the Escherichia coli genome by using the Streptococcus pyogenes type II CRISPR-Cas9 system to realize a variety of precise genome modifications, including gene deletion and insertion, with a highest efficiency of 100%, which was able to achieve simultaneous multigene editing of up to three targets. The system also demonstrated successful targeted chromosomal deletions in Tatumella citrea, another species of the Enterobacteriaceae, with highest efficiency of 100%.