Genome Engineering of Eubacterium limosum Using Expanded Genetic Tools and the CRISPR-Cas9 System
- Title
- Genome Engineering of Eubacterium limosum Using Expanded Genetic Tools and the CRISPR-Cas9 System
- Author
- 김동립
- Keywords
- Eubacterium limosum; acetogen; inducible promoter; CRISPR-Cas9; CRISPR interference; Wood-Ljungdahl pathway
- Issue Date
- 2019-09
- Publisher
- AMER CHEMICAL SOC
- Citation
- ACS SYNTHETIC BIOLOGY, v. 8, no. 9, Page. 2059-2068
- Abstract
- 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.
- URI
- https://pubs.acs.org/doi/10.1021/acssynbio.9b00150https://repository.hanyang.ac.kr/handle/20.500.11754/153570
- ISSN
- 2161-5063
- DOI
- 10.1021/acssynbio.9b00150
- Appears in Collections:
- COLLEGE OF ENGINEERING[S](공과대학) > MECHANICAL ENGINEERING(기계공학부) > Articles
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