CRISPR/Cas9 genome editing tool for glutamate bacillus.

Glutamate bacillus is an important amino acid production strain, its amino acid production of more than 4 million tons per year, in recent years has been widely used in the production of a variety of natural and non-natural products, its fermentation products are expected to reach a market value of 20.4 billion U.S. dollars by 2020.
the traditional industrial strains mainly rely on long-term physical and chemical mutagenic and screening to obtain, the genome level to achieve rapid and efficient rational editing is still the difficulty of glutamate barbacillus metabolic engineering transformation.
recently, the Systems and Synthetic Biotechnology Research Team of Tianjin Institute of Industrial Biotechnology of the Chinese Academy of Sciences developed the CRISPR/Cas9 genome editing tool of glutamate bacillus.
the study solved the problem that Cas9 toxicity and gRNA's own transcription terminator could not be terminated by reconstructing the expression box of Cas9 and gRNA, achieved efficient reverse screening based on CRISPR/Cas9 in glutamate bacillus;
in addition, the team developed CRISPR/Cas9 and RecT-mediated ssDNA recombination technology that can accurately introduce small-scale mutations and single-base mutations in the genome, with unit editing efficiency of more than 80% and two-site editing efficiency of up to 40%.
CRISPR/Cas9 genome editing tool of glutamate barbacillus has universality in the pattern strain glutamate bar ATCC 13032 and other polystrains of glutamate barbacillus, which can significantly improve the level of metabolic engineering transformation of glutamate barcobacillus and promote its production as an industrial chassis for a variety of bio-based chemicals and biofuels.
research published in Microb Cell Fact.
the research was supported by the National Natural Science Foundation of China, the key deployment project of the Chinese Academy of Sciences, and the Tianjin Special Branch Project.
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