Markerless chromosomal gene deletion in Clostridium beijerinckii using CRISPR/Cas9 system

Yi Wang; Zhong Tian Zhang; Seung Oh Seo; Kijoong Choi; Ting Lu; Yong Su Jin; Hans P. Blaschek

doi:10.1016/j.jbiotec.2015.02.005

Markerless chromosomal gene deletion in Clostridium beijerinckii using CRISPR/Cas9 system

Yi Wang, Zhong Tian Zhang, Seung Oh Seo, Kijoong Choi, Ting Lu, Yong Su Jin, Hans P. Blaschek

Biotechnology Institute

Research output: Contribution to journal › Article › peer-review

143 Scopus citations

Abstract

The anaerobic spore-forming, gram-positive, solventogenic clostridia are notorious for being difficult to genetically engineer. Based on CRISPR/Cas9 assisted homologous recombination, we demonstrated that clean markerless gene deletion from the chromosome can be easily achieved with a high efficiency through a single-step transformation in Clostridium beijerinckii NCIMB 8052, one of the most prominent strains for acetone, butanol and ethanol (ABE) production. This highly efficient genome engineering system can be further explored for multiplex genome engineering purposes. The protocols and principles developed in this study provided valuable references for genome engineering in other microorganisms lacking developed genetic engineering tools.

Original language	English (US)
Pages (from-to)	1-5
Number of pages	5
Journal	Journal of Biotechnology
Volume	200
DOIs	https://doi.org/10.1016/j.jbiotec.2015.02.005
State	Published - Apr 1 2015

Bibliographical note

Publisher Copyright:
© 2015 Elsevier B.V.

Keywords

Butanol
CRISPR/Cas9
Clostridium
Genome engineering
Homologous recombination

Publisher link

10.1016/j.jbiotec.2015.02.005

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abstract = "The anaerobic spore-forming, gram-positive, solventogenic clostridia are notorious for being difficult to genetically engineer. Based on CRISPR/Cas9 assisted homologous recombination, we demonstrated that clean markerless gene deletion from the chromosome can be easily achieved with a high efficiency through a single-step transformation in Clostridium beijerinckii NCIMB 8052, one of the most prominent strains for acetone, butanol and ethanol (ABE) production. This highly efficient genome engineering system can be further explored for multiplex genome engineering purposes. The protocols and principles developed in this study provided valuable references for genome engineering in other microorganisms lacking developed genetic engineering tools.",

keywords = "Butanol, CRISPR/Cas9, Clostridium, Genome engineering, Homologous recombination",

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AU - Wang, Yi

AU - Zhang, Zhong Tian

AU - Seo, Seung Oh

AU - Choi, Kijoong

AU - Lu, Ting

AU - Jin, Yong Su

AU - Blaschek, Hans P.

PY - 2015/4/1

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AB - The anaerobic spore-forming, gram-positive, solventogenic clostridia are notorious for being difficult to genetically engineer. Based on CRISPR/Cas9 assisted homologous recombination, we demonstrated that clean markerless gene deletion from the chromosome can be easily achieved with a high efficiency through a single-step transformation in Clostridium beijerinckii NCIMB 8052, one of the most prominent strains for acetone, butanol and ethanol (ABE) production. This highly efficient genome engineering system can be further explored for multiplex genome engineering purposes. The protocols and principles developed in this study provided valuable references for genome engineering in other microorganisms lacking developed genetic engineering tools.

KW - Butanol

KW - CRISPR/Cas9

KW - Clostridium

KW - Genome engineering

KW - Homologous recombination

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Markerless chromosomal gene deletion in Clostridium beijerinckii using CRISPR/Cas9 system

Abstract

Bibliographical note

Keywords

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