• Journal of biotechnology · Apr 2015

    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, and Hans P Blaschek.
    • Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
    • J. Biotechnol. 2015 Apr 20; 200: 1-5.

    AbstractThe 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. Copyright © 2015 Elsevier B.V. All rights reserved.

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