Jiang Yu, Chen Biao, Duan Chunlan, Sun Bingbing, Yang Junjie, Yang Sheng
Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China Shanghai Research Center of Industrial Biotechnology, Shanghai, China.
Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
Appl Environ Microbiol. 2015 Apr;81(7):2506-14. doi: 10.1128/AEM.04023-14. Epub 2015 Jan 30.
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%.
构建具有工业用途的微生物需要一种高效的全基因组规模编辑工具。我们描述了一种靶向、连续多基因编辑策略，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，以实现多种精确的基因组修饰，包括基因缺失和插入，最高效率可达100%，能够实现多达三个靶点的同时多基因编辑。该系统还在肠杆菌科的另一个物种——柠檬塔特姆菌中成功实现了靶向染色体缺失，最高效率为100%。
