Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid
Botulinum neurotoxins (BoNTs), produced by the spore-forming bacterium Clostridium botulinum, cause botulism, a rare but fatal illness affecting humans and animals. Despite causing a life-threatening disease, BoNT is a multipurpose therapeutic. Nevertheless, as the most potent natural toxin, BoNT is...
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ftpubmed:oai:pubmedcentral.nih.gov:8810926 2023-05-15T15:41:32+02:00 Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid Nowakowska, Maria B. Selby, Katja Przykopanski, Adina Krüger, Maren Krez, Nadja Dorner, Brigitte G. Dorner, Martin B. Jin, Rongsheng Minton, Nigel P. Rummel, Andreas Lindström, Miia 2022-02-02 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8810926/ http://www.ncbi.nlm.nih.gov/pubmed/35110559 https://doi.org/10.1038/s41598-022-05008-1 en eng Nature Publishing Group UK http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8810926/ http://www.ncbi.nlm.nih.gov/pubmed/35110559 http://dx.doi.org/10.1038/s41598-022-05008-1 © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . CC-BY Sci Rep Article Text 2022 ftpubmed https://doi.org/10.1038/s41598-022-05008-1 2022-02-13T01:34:42Z Botulinum neurotoxins (BoNTs), produced by the spore-forming bacterium Clostridium botulinum, cause botulism, a rare but fatal illness affecting humans and animals. Despite causing a life-threatening disease, BoNT is a multipurpose therapeutic. Nevertheless, as the most potent natural toxin, BoNT is classified as a Select Agent in the US, placing C. botulinum research under stringent governmental regulations. The extreme toxicity of BoNT, its impact on public safety, and its diverse therapeutic applications urge to devise safe solutions to expand C. botulinum research. Accordingly, we exploited CRISPR/Cas9-mediated genome editing to introduce inactivating point mutations into chromosomal bont/e gene of C. botulinum Beluga E. The resulting Beluga Ei strain displays unchanged physiology and produces inactive BoNT (BoNT/Ei) recognized in serological assays, but lacking biological activity detectable ex- and in vivo. Neither native single-chain, nor trypsinized di-chain form of BoNT/Ei show in vivo toxicity, even if isolated from Beluga Ei sub-cultured for 25 generations. Beluga Ei strain constitutes a safe alternative for the BoNT research necessary for public health risk management, the development of food preservation strategies, understanding toxinogenesis, and for structural BoNT studies. The example of Beluga Ei generation serves as template for future development of C. botulinum producing different inactive BoNT serotypes. Text Beluga Beluga* PubMed Central (PMC) Scientific Reports 12 1 |
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Article Nowakowska, Maria B. Selby, Katja Przykopanski, Adina Krüger, Maren Krez, Nadja Dorner, Brigitte G. Dorner, Martin B. Jin, Rongsheng Minton, Nigel P. Rummel, Andreas Lindström, Miia Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid |
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Botulinum neurotoxins (BoNTs), produced by the spore-forming bacterium Clostridium botulinum, cause botulism, a rare but fatal illness affecting humans and animals. Despite causing a life-threatening disease, BoNT is a multipurpose therapeutic. Nevertheless, as the most potent natural toxin, BoNT is classified as a Select Agent in the US, placing C. botulinum research under stringent governmental regulations. The extreme toxicity of BoNT, its impact on public safety, and its diverse therapeutic applications urge to devise safe solutions to expand C. botulinum research. Accordingly, we exploited CRISPR/Cas9-mediated genome editing to introduce inactivating point mutations into chromosomal bont/e gene of C. botulinum Beluga E. The resulting Beluga Ei strain displays unchanged physiology and produces inactive BoNT (BoNT/Ei) recognized in serological assays, but lacking biological activity detectable ex- and in vivo. Neither native single-chain, nor trypsinized di-chain form of BoNT/Ei show in vivo toxicity, even if isolated from Beluga Ei sub-cultured for 25 generations. Beluga Ei strain constitutes a safe alternative for the BoNT research necessary for public health risk management, the development of food preservation strategies, understanding toxinogenesis, and for structural BoNT studies. The example of Beluga Ei generation serves as template for future development of C. botulinum producing different inactive BoNT serotypes. |
format |
Text |
author |
Nowakowska, Maria B. Selby, Katja Przykopanski, Adina Krüger, Maren Krez, Nadja Dorner, Brigitte G. Dorner, Martin B. Jin, Rongsheng Minton, Nigel P. Rummel, Andreas Lindström, Miia |
author_facet |
Nowakowska, Maria B. Selby, Katja Przykopanski, Adina Krüger, Maren Krez, Nadja Dorner, Brigitte G. Dorner, Martin B. Jin, Rongsheng Minton, Nigel P. Rummel, Andreas Lindström, Miia |
author_sort |
Nowakowska, Maria B. |
title |
Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid |
title_short |
Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid |
title_full |
Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid |
title_fullStr |
Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid |
title_full_unstemmed |
Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid |
title_sort |
construction and validation of safe clostridium botulinum group ii surrogate strain producing inactive botulinum neurotoxin type e toxoid |
publisher |
Nature Publishing Group UK |
publishDate |
2022 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8810926/ http://www.ncbi.nlm.nih.gov/pubmed/35110559 https://doi.org/10.1038/s41598-022-05008-1 |
genre |
Beluga Beluga* |
genre_facet |
Beluga Beluga* |
op_source |
Sci Rep |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8810926/ http://www.ncbi.nlm.nih.gov/pubmed/35110559 http://dx.doi.org/10.1038/s41598-022-05008-1 |
op_rights |
© The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
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CC-BY |
op_doi |
https://doi.org/10.1038/s41598-022-05008-1 |
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