A Strategy for the Rapid Development of a Safe Vibrio cholerae Candidate Vaccine Strain

Approximately 1/6 of humanity is at high risk of experiencing cholera epidemics. The development of effective and safe vaccines against Vibrio cholerae, the primary cause of cholera, is part of the public health measures to prevent cholera epidemics. Natural nontoxigenic ...

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Autores principales:	Dmitry S. Karpov, Anna V. Goncharenko, Evgenii V. Usachev, Daria V. Vasina, Elizaveta V. Divisenko, Yaroslava M. Chalenko, Andrei A. Pochtovyi, Roman S. Ovchinnikov, Valentin V. Makarov, Sergei M. Yudin, Artem P. Tkachuk, Vladimir A. Gushchin
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Vibrio cholerae</i> genome engineering synthetic reporter operon amilCP candidate vaccine strain Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/796c770d9b3442c18ad01745fc78b4dc
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spelling	oai:doaj.org-article:796c770d9b3442c18ad01745fc78b4dc2021-11-11T17:07:33ZA Strategy for the Rapid Development of a Safe <i>Vibrio cholerae</i> Candidate Vaccine Strain10.3390/ijms2221116571422-00671661-6596https://doaj.org/article/796c770d9b3442c18ad01745fc78b4dc2021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11657https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Approximately 1/6 of humanity is at high risk of experiencing cholera epidemics. The development of effective and safe vaccines against <i>Vibrio cholerae,</i> the primary cause of cholera, is part of the public health measures to prevent cholera epidemics. Natural nontoxigenic <i>V. cholerae</i> isolates represent a source of new genetically improved and relatively safe vaccine strains. However, the genomic engineering of wild-type <i>V. cholerae</i> strains is difficult, and these strains are genetically unstable due to their high homologous recombination activity. We comprehensively characterized two <i>V. cholerae</i> isolates using genome sequencing, bioinformatic analysis, and microscopic, physiological, and biochemical tests. Genetic constructs were Gibson assembled and electrotransformed into <i>V. cholerae</i>. Bacterial colonies were assessed using standard microbiological and immunological techniques. As a result, we created a synthetic chromoprotein-expressing reporter operon. This operon was used to improve the <i>V. cholerae</i> genome engineering approach and monitor the stability of the genetic constructs. Finally, we created a stable candidate <i>V. cholerae</i> vaccine strain bearing a <i>recA</i> deletion and expressing the β-subunit of cholera toxin. Thus, we developed a strategy for the rapid creation of genetically stable and relatively safe candidate vaccine strains. This strategy can be applied not only to <i>V. cholerae</i> but also to other important human bacterial pathogens.Dmitry S. KarpovAnna V. GoncharenkoEvgenii V. UsachevDaria V. VasinaElizaveta V. DivisenkoYaroslava M. ChalenkoAndrei A. PochtovyiRoman S. OvchinnikovValentin V. MakarovSergei M. YudinArtem P. TkachukVladimir A. GushchinMDPI AGarticle<i>Vibrio cholerae</i>genome engineeringsynthetic reporter operonamilCPcandidate vaccine strainBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11657, p 11657 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Vibrio cholerae</i> genome engineering synthetic reporter operon amilCP candidate vaccine strain Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	<i>Vibrio cholerae</i> genome engineering synthetic reporter operon amilCP candidate vaccine strain Biology (General) QH301-705.5 Chemistry QD1-999 Dmitry S. Karpov Anna V. Goncharenko Evgenii V. Usachev Daria V. Vasina Elizaveta V. Divisenko Yaroslava M. Chalenko Andrei A. Pochtovyi Roman S. Ovchinnikov Valentin V. Makarov Sergei M. Yudin Artem P. Tkachuk Vladimir A. Gushchin A Strategy for the Rapid Development of a Safe <i>Vibrio cholerae</i> Candidate Vaccine Strain
description	Approximately 1/6 of humanity is at high risk of experiencing cholera epidemics. The development of effective and safe vaccines against <i>Vibrio cholerae,</i> the primary cause of cholera, is part of the public health measures to prevent cholera epidemics. Natural nontoxigenic <i>V. cholerae</i> isolates represent a source of new genetically improved and relatively safe vaccine strains. However, the genomic engineering of wild-type <i>V. cholerae</i> strains is difficult, and these strains are genetically unstable due to their high homologous recombination activity. We comprehensively characterized two <i>V. cholerae</i> isolates using genome sequencing, bioinformatic analysis, and microscopic, physiological, and biochemical tests. Genetic constructs were Gibson assembled and electrotransformed into <i>V. cholerae</i>. Bacterial colonies were assessed using standard microbiological and immunological techniques. As a result, we created a synthetic chromoprotein-expressing reporter operon. This operon was used to improve the <i>V. cholerae</i> genome engineering approach and monitor the stability of the genetic constructs. Finally, we created a stable candidate <i>V. cholerae</i> vaccine strain bearing a <i>recA</i> deletion and expressing the β-subunit of cholera toxin. Thus, we developed a strategy for the rapid creation of genetically stable and relatively safe candidate vaccine strains. This strategy can be applied not only to <i>V. cholerae</i> but also to other important human bacterial pathogens.
format	article
author	Dmitry S. Karpov Anna V. Goncharenko Evgenii V. Usachev Daria V. Vasina Elizaveta V. Divisenko Yaroslava M. Chalenko Andrei A. Pochtovyi Roman S. Ovchinnikov Valentin V. Makarov Sergei M. Yudin Artem P. Tkachuk Vladimir A. Gushchin
author_facet	Dmitry S. Karpov Anna V. Goncharenko Evgenii V. Usachev Daria V. Vasina Elizaveta V. Divisenko Yaroslava M. Chalenko Andrei A. Pochtovyi Roman S. Ovchinnikov Valentin V. Makarov Sergei M. Yudin Artem P. Tkachuk Vladimir A. Gushchin
author_sort	Dmitry S. Karpov
title	A Strategy for the Rapid Development of a Safe <i>Vibrio cholerae</i> Candidate Vaccine Strain
title_short	A Strategy for the Rapid Development of a Safe <i>Vibrio cholerae</i> Candidate Vaccine Strain
title_full	A Strategy for the Rapid Development of a Safe <i>Vibrio cholerae</i> Candidate Vaccine Strain
title_fullStr	A Strategy for the Rapid Development of a Safe <i>Vibrio cholerae</i> Candidate Vaccine Strain
title_full_unstemmed	A Strategy for the Rapid Development of a Safe <i>Vibrio cholerae</i> Candidate Vaccine Strain
title_sort	strategy for the rapid development of a safe <i>vibrio cholerae</i> candidate vaccine strain
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/796c770d9b3442c18ad01745fc78b4dc
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A Strategy for the Rapid Development of a Safe <i>Vibrio cholerae</i> Candidate Vaccine Strain

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