Markerless chromosomal gene deletion in Clostridium beijerinckii using CRISPR/Cas9 system
Open accessAbstract
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.
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The CRISPR/Cas9 system from Streptococcus pyogenes was successfully applied to various clostridia to deliver one-step, markerless, and highly efficient gene editing.
“Recently, the CRISPR/Cas9 system from Streptococcus pyogenes was successfully applied to various clostridia (Wang et al., 2015; Xu et al., 2015) to deliver one-step, markerless, and highly efficient gene editing.”
From Gas Fermentation—A Flexible Platform for Commercial Scale Production of Low-Carbon-Fuels and Chemicals from Waste and Renewable Feedstocks (2016) · cites this paper as Wang et al., 2015
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References 29 · Cited by 161
- Authors
- Wang, Y., Zhang, Z.-T., Seo, S.-O., Choi, K., Lu, T., Jin, Y.-S., & Blaschek, H. P.
- Affiliations
- University of Illinois Urbana-Champaign (US)
- Corresponding author
- Blaschek, Hans P.
- Venue
- Journal of Biotechnology
- Publisher
- Elsevier BV
- Published
- 2015-04
- Volume
- 200
- Pages
- 1-5
- ISSN
- 0168-1656 (ISSN-L)
- OpenAlex
- W2066991387
- Cited by
- 161
- References
- 29
- Citations by year
- 2026: 5 · 2025: 2 · 2024: 1 · 2023: 10 · 2022: 11 · 2021: 8 · 2020: 10 · 2019: 31 · 2018: 16 · 2017: 22 · 2016: 34 · 2015: 10
- FWCI
- 11.86
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.elsevier.com/tdm/userlicense/1.0/ (tdm)
- www.elsevier.com/legal/tdmrep-license (tdm)
- www.elsevier.com/open-access/userlicense/1.0/ (am, 455 d)
- doi.org/10.15223/policy-017 (stm-asf)
- doi.org/10.15223/policy-037 (stm-asf)
- doi.org/10.15223/policy-012 (stm-asf)
- doi.org/10.15223/policy-029 (stm-asf)
- doi.org/10.15223/policy-004 (stm-asf)
- Locations
- Journal of Biotechnology (acceptedVersion)
- PubMed (publishedVersion)
- OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
- Funders
- U.S. Department of Energy (2011-01219)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- markerless gene deletion, Clostridium beijerinckii, CRISPR/Cas9, homologous recombination, genome editing, solventogenic clostridia
- MeSH terms
- Chromosomes, Bacterial, Genes, Bacterial, Genetic Engineering, Genome, Bacterial, Gene Deletion, Clostridium beijerinckii, Clustered Regularly Interspaced Short Palindromic Repeats
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