Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 81, Issue 7, Pages 2506-2514Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.04023-14
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Funding
- National Basic Research Program of China [2012CB721105, 2014CB745101]
- National High Technology Research and Development Program of China [2012AA02A704]
- Knowledge Innovation Program [KSZD-EW-Z-016-1, KSZD-EW-Z-019]
- Science and Technology Service Network Initiative of the Chinese Academy of Sciences [KFJ-EW-STS-030]
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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%.
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