Genome Engineering of Eubacterium limosum Using Expanded Genetic Tools and the CRISPR-Cas9 System

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Eubacterium limosum is one of the important bacteria in C-1 feedstock utilization as well as in human gut microbiota. Although E. limosum has recently garnered much attention and investigation on a genome-wide scale, a bottleneck for systematic engineering in E. limosum is the lack of available genetic tools and an efficient genome editing platform. To overcome this limitation, we here report expanded genetic tools and the CRISPR-Cas9 system. We have developed an inducible promoter system that enables implementation of the CRISPR-Cas9 system to precisely manipulate target genes of the Wood-Ljungdahl pathway with 100% efficiency. Furthermore, we exploited the effectiveness of CRISPR interference to reduce the expression of target genes, exhibiting substantial repression of several genes in the Wood-Ljungdahl pathway and fructose-PTS system. These expanded genetic tools and CRISPR-Cas9 system comprise powerful and widely applicable genetic tools to accelerate functional genomic study and genome engineering in E. limosum.
Publisher
AMER CHEMICAL SOC
Issue Date
2019-09
Language
English
Article Type
Article
Citation

ACS SYNTHETIC BIOLOGY, v.8, no.9, pp.2059 - 2068

ISSN
2161-5063
DOI
10.1021/acssynbio.9b00150
URI
http://hdl.handle.net/10203/267993
Appears in Collection
BS-Journal Papers(저널논문)
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