For the first time in E. coli, CRISPR/Cas9 technology was used to achieve high-efficiency editing of four genes at the same time.

The genome editing technology of industrial microorganisms is the research foundation and hot spot of microbial metabolic engineering, and the establishment of CRISPR/Cas9 system has greatly promoted the development of genome editing technology.
however, the problem of crispR/Cas9 multigene simultaneous editing efficiency still exists in the model bio-E. coli, and the lack of effective genome editing methods for Ralstonia eutropha, which has great potential for industrial application, severely restricts the in-depth study of Roche's real bacteria.
recently, the research team of microbial metabolic engineering and synthetic biotechnology led by Zhang Xueli, a researcher of Tianjin Institute of Industrial Biotechnology of the Chinese Academy of Sciences, led by Bi Changxuan, used CRISPR/Cas9 technology for the first time in E. coli to achieve high-efficiency editing of four genes, with two genes, three genes and four genes editing efficiency at the same time reaching 100%, 90% and 40% respectively.
results have been published in Biotechnology Journal, Tianjin Institute of Technology Assistant Researcher Zhao Dongdong and master's student Feng Xu as the first author of the paper. In addition, the team has developed an efficient genome editing method in Roche's true bacteria,
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increased the efficiency of strain conversion by three orders of magnitude by knocking out the restrictive endoenzyme gene in Roche's gene-restriction modification system, and achieved gene editing efficiency of 78.3-100% using CRISPR-Cas9-mediated homologous recombination (HR).
this technology overcomes the technical bottleneck of Roche's genome editing, provides a new method of genetic modification of the strain, and lays the foundation for the industrialization application of the bacteria.
results have been published in the journal Biotechnology for Biofuels, with Xiong Bin, a doctoral student at Tianjin Institute of Technology, as the first author of the paper.
research supported by the National "863" (2015AA02020202), the Key Projects of the Tianjin Science and Technology Support Program (14ZCZDSY00067) and the Key Deployment Project of the Chinese Academy of Sciences (ZDRW-ZS-2016-3).
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