<named-content content-type="genus-species">Enterococcus faecalis</named-content> CRISPR-Cas Is a Robust Barrier to Conjugative Antibiotic Resistance Dissemination in the Murine Intestine

ABSTRACT CRISPR-Cas systems are barriers to horizontal gene transfer (HGT) in bacteria. Little is known about CRISPR-Cas interactions with conjugative plasmids, and studies investigating CRISPR-Cas/plasmid interactions in in vivo models relevant to infectious disease are lacking. These are significa...

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Autores principales:	Valerie J. Price, Sara W. McBride, Karthik Hullahalli, Anushila Chatterjee, Breck A. Duerkop, Kelli L. Palmer
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	CRISPR-Cas Enterococcus faecalis antibiotic resistance intestinal colonization plasmids Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/9306573beb6b4c9cbff038b7015839c5
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spelling	oai:doaj.org-article:9306573beb6b4c9cbff038b7015839c52021-11-15T15:22:27Z<named-content content-type="genus-species">Enterococcus faecalis</named-content> CRISPR-Cas Is a Robust Barrier to Conjugative Antibiotic Resistance Dissemination in the Murine Intestine10.1128/mSphere.00464-192379-5042https://doaj.org/article/9306573beb6b4c9cbff038b7015839c52019-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00464-19https://doaj.org/toc/2379-5042ABSTRACT CRISPR-Cas systems are barriers to horizontal gene transfer (HGT) in bacteria. Little is known about CRISPR-Cas interactions with conjugative plasmids, and studies investigating CRISPR-Cas/plasmid interactions in in vivo models relevant to infectious disease are lacking. These are significant gaps in knowledge because conjugative plasmids disseminate antibiotic resistance genes among pathogens in vivo, and it is essential to identify strategies to reduce the spread of these elements. We use enterococci as models to understand the interactions of CRISPR-Cas with conjugative plasmids. Enterococcus faecalis is a native colonizer of the mammalian intestine and harbors pheromone-responsive plasmids (PRPs). PRPs mediate inter- and intraspecies transfer of antibiotic resistance genes. We assessed E. faecalis CRISPR-Cas anti-PRP activity in the mouse intestine and under different in vitro conditions. We observed striking differences in CRISPR-Cas efficiency in vitro versus in vivo. With few exceptions, CRISPR-Cas blocked intestinal PRP dissemination, while in vitro, the PRP frequently escaped CRISPR-Cas defense. Our results further the understanding of CRISPR-Cas biology by demonstrating that standard in vitro experiments do not adequately model the in vivo antiplasmid activity of CRISPR-Cas. Additionally, our work identifies several variables that impact the apparent in vitro antiplasmid activity of CRISPR-Cas, including planktonic versus biofilm settings, different donor-to-recipient ratios, production of a plasmid-encoded bacteriocin, and the time point at which matings are sampled. Our results are clinically significant because they demonstrate that barriers to HGT encoded by normal (healthy) human microbiota can have significant impacts on in vivo antibiotic resistance dissemination. IMPORTANCE CRISPR-Cas is a type of immune system in bacteria that is hypothesized to be a natural impediment to the spread of antibiotic resistance genes. In this study, we directly assessed the impact of CRISPR-Cas on antibiotic resistance dissemination in the mammalian intestine and under different in vitro conditions. We observed a robust effect of CRISPR-Cas on in vivo but not in vitro dissemination of antibiotic resistance plasmids in the native mammalian intestinal colonizer Enterococcus faecalis. We conclude that standard in vitro experiments currently do not appropriately model the in vivo conditions where antibiotic resistance dissemination occurs between E. faecalis strains in the intestine. Moreover, our results demonstrate that CRISPR-Cas present in native members of the mammalian intestinal microbiota can block the spread of antibiotic resistance plasmids.Valerie J. PriceSara W. McBrideKarthik HullahalliAnushila ChatterjeeBreck A. DuerkopKelli L. PalmerAmerican Society for MicrobiologyarticleCRISPR-CasEnterococcus faecalisantibiotic resistanceintestinal colonizationplasmidsMicrobiologyQR1-502ENmSphere, Vol 4, Iss 4 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	CRISPR-Cas Enterococcus faecalis antibiotic resistance intestinal colonization plasmids Microbiology QR1-502
spellingShingle	CRISPR-Cas Enterococcus faecalis antibiotic resistance intestinal colonization plasmids Microbiology QR1-502 Valerie J. Price Sara W. McBride Karthik Hullahalli Anushila Chatterjee Breck A. Duerkop Kelli L. Palmer <named-content content-type="genus-species">Enterococcus faecalis</named-content> CRISPR-Cas Is a Robust Barrier to Conjugative Antibiotic Resistance Dissemination in the Murine Intestine
description	ABSTRACT CRISPR-Cas systems are barriers to horizontal gene transfer (HGT) in bacteria. Little is known about CRISPR-Cas interactions with conjugative plasmids, and studies investigating CRISPR-Cas/plasmid interactions in in vivo models relevant to infectious disease are lacking. These are significant gaps in knowledge because conjugative plasmids disseminate antibiotic resistance genes among pathogens in vivo, and it is essential to identify strategies to reduce the spread of these elements. We use enterococci as models to understand the interactions of CRISPR-Cas with conjugative plasmids. Enterococcus faecalis is a native colonizer of the mammalian intestine and harbors pheromone-responsive plasmids (PRPs). PRPs mediate inter- and intraspecies transfer of antibiotic resistance genes. We assessed E. faecalis CRISPR-Cas anti-PRP activity in the mouse intestine and under different in vitro conditions. We observed striking differences in CRISPR-Cas efficiency in vitro versus in vivo. With few exceptions, CRISPR-Cas blocked intestinal PRP dissemination, while in vitro, the PRP frequently escaped CRISPR-Cas defense. Our results further the understanding of CRISPR-Cas biology by demonstrating that standard in vitro experiments do not adequately model the in vivo antiplasmid activity of CRISPR-Cas. Additionally, our work identifies several variables that impact the apparent in vitro antiplasmid activity of CRISPR-Cas, including planktonic versus biofilm settings, different donor-to-recipient ratios, production of a plasmid-encoded bacteriocin, and the time point at which matings are sampled. Our results are clinically significant because they demonstrate that barriers to HGT encoded by normal (healthy) human microbiota can have significant impacts on in vivo antibiotic resistance dissemination. IMPORTANCE CRISPR-Cas is a type of immune system in bacteria that is hypothesized to be a natural impediment to the spread of antibiotic resistance genes. In this study, we directly assessed the impact of CRISPR-Cas on antibiotic resistance dissemination in the mammalian intestine and under different in vitro conditions. We observed a robust effect of CRISPR-Cas on in vivo but not in vitro dissemination of antibiotic resistance plasmids in the native mammalian intestinal colonizer Enterococcus faecalis. We conclude that standard in vitro experiments currently do not appropriately model the in vivo conditions where antibiotic resistance dissemination occurs between E. faecalis strains in the intestine. Moreover, our results demonstrate that CRISPR-Cas present in native members of the mammalian intestinal microbiota can block the spread of antibiotic resistance plasmids.
format	article
author	Valerie J. Price Sara W. McBride Karthik Hullahalli Anushila Chatterjee Breck A. Duerkop Kelli L. Palmer
author_facet	Valerie J. Price Sara W. McBride Karthik Hullahalli Anushila Chatterjee Breck A. Duerkop Kelli L. Palmer
author_sort	Valerie J. Price
title	<named-content content-type="genus-species">Enterococcus faecalis</named-content> CRISPR-Cas Is a Robust Barrier to Conjugative Antibiotic Resistance Dissemination in the Murine Intestine
title_short	<named-content content-type="genus-species">Enterococcus faecalis</named-content> CRISPR-Cas Is a Robust Barrier to Conjugative Antibiotic Resistance Dissemination in the Murine Intestine
title_full	<named-content content-type="genus-species">Enterococcus faecalis</named-content> CRISPR-Cas Is a Robust Barrier to Conjugative Antibiotic Resistance Dissemination in the Murine Intestine
title_fullStr	<named-content content-type="genus-species">Enterococcus faecalis</named-content> CRISPR-Cas Is a Robust Barrier to Conjugative Antibiotic Resistance Dissemination in the Murine Intestine
title_full_unstemmed	<named-content content-type="genus-species">Enterococcus faecalis</named-content> CRISPR-Cas Is a Robust Barrier to Conjugative Antibiotic Resistance Dissemination in the Murine Intestine
title_sort	<named-content content-type="genus-species">enterococcus faecalis</named-content> crispr-cas is a robust barrier to conjugative antibiotic resistance dissemination in the murine intestine
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/9306573beb6b4c9cbff038b7015839c5
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<named-content content-type="genus-species">Enterococcus faecalis</named-content> CRISPR-Cas Is a Robust Barrier to Conjugative Antibiotic Resistance Dissemination in the Murine Intestine

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