Construction and validation of safe Clostridium botulinum Group II surrogate strain producing inactive botulinum neurotoxin type E toxoid.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
02 02 2022
02 02 2022
Historique:
received:
18
10
2021
accepted:
24
12
2021
entrez:
3
2
2022
pubmed:
4
2
2022
medline:
11
3
2022
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 35110559
doi: 10.1038/s41598-022-05008-1
pii: 10.1038/s41598-022-05008-1
pmc: PMC8810926
doi:
Substances chimiques
CRISPR-Associated Protein 9
EC 3.1.-
Botulinum Toxins
EC 3.4.24.69
botulinum toxin type E
T579M564JY
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1790Subventions
Organisme : NIAID NIH HHS
ID : R01 AI125704
Pays : United States
Informations de copyright
© 2022. The Author(s).
Références
J Infect Dis. 2014 Jan 15;209(2):192-202
pubmed: 24106295
Toxins (Basel). 2019 Dec 07;11(12):
pubmed: 31817843
J Biol Chem. 1992 Nov 25;267(33):23479-83
pubmed: 1429690
Int J Food Microbiol. 2006 Apr 15;108(1):92-104
pubmed: 16480785
Fed Regist. ;82(12):6278-94
pubmed: 28106357
J Appl Microbiol. 2006 Sep;101(3):556-70
pubmed: 16907806
Toxicon. 2001 Aug;39(8):1151-9
pubmed: 11306125
Mol Biol Evol. 2016 Jul;33(7):1870-4
pubmed: 27004904
Environ Microbiol. 2017 Oct;19(10):4287-4300
pubmed: 28809452
Nucleic Acids Res. 1994 Nov 11;22(22):4673-80
pubmed: 7984417
Trends Cell Biol. 1994 May;4(5):179-85
pubmed: 14731646
Physiol Rev. 2000 Apr;80(2):717-66
pubmed: 10747206
Biochemistry. 2004 Jun 1;43(21):6637-44
pubmed: 15157097
Sci Rep. 2019 Apr 2;9(1):5531
pubmed: 30940836
J Bacteriol. 1956 Oct;72(4):455-60
pubmed: 13366946
Nat Commun. 2017 Aug 03;8:14130
pubmed: 28770820
Biochem Biophys Res Commun. 2001 Nov 16;288(5):1231-7
pubmed: 11700044
Biochemistry. 2000 Mar 7;39(9):2399-405
pubmed: 10694409
Clin Infect Dis. 2005 Oct 15;41(8):1167-73
pubmed: 16163636
J Neurochem. 2009 Jun;109(6):1584-95
pubmed: 19457120
Front Microbiol. 2021 Jan 27;12:617269
pubmed: 33584620
Biochemistry. 2006 Jul 25;45(29):8903-11
pubmed: 16846233
J Physiol Paris. 1995;89(1):43-50
pubmed: 7581298
J Mol Biol. 2009 Feb 13;386(1):233-45
pubmed: 19118561
Curr Top Microbiol Immunol. 2013;364:139-57
pubmed: 23239352
Int J Food Microbiol. 1997 Jun 17;37(1):63-72
pubmed: 9237123
Appl Environ Microbiol. 2010 Jul;76(13):4448-60
pubmed: 20453132
Res Microbiol. 2015 May;166(4):303-17
pubmed: 25445012
J Mol Biol. 1999 Sep 3;291(5):1091-104
pubmed: 10518945
Nat Struct Biol. 1998 Oct;5(10):898-902
pubmed: 9783750
Nat Struct Biol. 2000 Aug;7(8):693-9
pubmed: 10932256
Lett Appl Microbiol. 1997 Feb;24(2):95-100
pubmed: 9081311
Biochemistry. 2002 Feb 12;41(6):1717-23
pubmed: 11827515
Science. 2012 Feb 24;335(6071):977-81
pubmed: 22363010
Mol Microbiol. 2002 Oct;46(2):439-52
pubmed: 12406220
Infect Immun. 2003 Mar;71(3):1147-54
pubmed: 12595426
Muscle Nerve. 1998 Jun;21(6):701-10
pubmed: 9585323
BMJ Open Sci. 2020 Jul 20;4(1):e100115
pubmed: 34095516
J Appl Microbiol. 1998 Jan;84(1):5-17
pubmed: 15244052
Toxins (Basel). 2015 Nov 25;7(12):4895-905
pubmed: 26610569
ACS Synth Biol. 2019 Jun 21;8(6):1379-1390
pubmed: 31181894
Nat Rev Neurol. 2010 Nov;6(11):624-36
pubmed: 21045798
Anal Chem. 2015 Apr 7;87(7):3911-7
pubmed: 25731972
Appl Environ Microbiol. 2016 May 02;82(10):3100-3108
pubmed: 26994073
Nature. 2004 Dec 16;432(7019):925-9
pubmed: 15592454
Food Microbiol. 2011 Apr;28(2):183-91
pubmed: 21315972
Sci Rep. 2019 May 31;9(1):8123
pubmed: 31148548
Toxins (Basel). 2015 Nov 26;7(12):5011-34
pubmed: 26703727
Sci Rep. 2018 Mar 14;8(1):4518
pubmed: 29540745
J Microbiol Methods. 2015 Jan;108:83-91
pubmed: 25433276
Nat Rev Microbiol. 2014 Aug;12(8):535-49
pubmed: 24975322
FEBS J. 2011 Dec;278(23):4467-85
pubmed: 21592305