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

ACS Synthetic Biology
Jongoh ShinByung-Kwan Cho

Abstract

Eubacterium limosum is one of the important bacteria in C1 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.

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Citations

Mar 29, 2020·Frontiers in Microbiology·Seulgi KangByung-Kwan Cho
Oct 21, 2020·International Journal of Molecular Sciences·Sangrak JinByung-Kwan Cho
Apr 22, 2020·Current Opinion in Biotechnology·Michael Köpke, Séan D Simpson
Feb 18, 2021·FEMS Microbiology Reviews·Barbara BourgadeM Ahsanul Islam
Apr 20, 2021·Annual Review of Chemical and Biomolecular Engineering·Nick FacklerMichael Köpke
Jul 2, 2020·ACS Synthetic Biology·Peng-Fei XiaBastian Molitor
Nov 30, 2021·Microbial Biotechnology·Jamin C WoodBernardino Virdis

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