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Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of Yersinia pestis Pulmonary Infection

Pneumonic plague, caused by Yersinia pestis, is a rapidly progressing lethal infection. The various phases of pneumonic plague are yet to be fully understood. A well-established way to address the pathology of infectious diseases in general, and pneumonic plague in particular, is to conduct concomit...

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Autores principales: Israeli, Ofir, Cohen-Gihon, Inbar, Aftalion, Moshe, Gur, David, Vagima, Yaron, Zauberman, Ayelet, Levy, Yinon, Zvi, Anat, Chitlaru, Theodor, Mamroud, Emanuelle, Tidhar, Avital
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8539884/
https://www.ncbi.nlm.nih.gov/pubmed/34683487
http://dx.doi.org/10.3390/microorganisms9102166
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author Israeli, Ofir
Cohen-Gihon, Inbar
Aftalion, Moshe
Gur, David
Vagima, Yaron
Zauberman, Ayelet
Levy, Yinon
Zvi, Anat
Chitlaru, Theodor
Mamroud, Emanuelle
Tidhar, Avital
author_facet Israeli, Ofir
Cohen-Gihon, Inbar
Aftalion, Moshe
Gur, David
Vagima, Yaron
Zauberman, Ayelet
Levy, Yinon
Zvi, Anat
Chitlaru, Theodor
Mamroud, Emanuelle
Tidhar, Avital
author_sort Israeli, Ofir
collection PubMed
description Pneumonic plague, caused by Yersinia pestis, is a rapidly progressing lethal infection. The various phases of pneumonic plague are yet to be fully understood. A well-established way to address the pathology of infectious diseases in general, and pneumonic plague in particular, is to conduct concomitant transcriptomic analysis of the bacteria and the host. The analysis of dual RNA by RNA sequencing technology is challenging, due the difficulties of extracting bacterial RNA, which is overwhelmingly outnumbered by the host RNA, especially at the critical early time points post-infection (prior to 48 h). Here, we describe a novel technique that employed the infusion of an RNA preserving reagent (RNAlater) into the lungs of the animals, through the trachea, under deep anesthesia. This method enabled the isolation of stable dual mRNA from the lungs of mice infected with Y. pestis, as early as 24 h post-infection. The RNA was used for transcriptomic analysis, which provided a comprehensive gene expression profile of both the host and the pathogen.
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spelling pubmed-85398842021-10-24 Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of Yersinia pestis Pulmonary Infection Israeli, Ofir Cohen-Gihon, Inbar Aftalion, Moshe Gur, David Vagima, Yaron Zauberman, Ayelet Levy, Yinon Zvi, Anat Chitlaru, Theodor Mamroud, Emanuelle Tidhar, Avital Microorganisms Communication Pneumonic plague, caused by Yersinia pestis, is a rapidly progressing lethal infection. The various phases of pneumonic plague are yet to be fully understood. A well-established way to address the pathology of infectious diseases in general, and pneumonic plague in particular, is to conduct concomitant transcriptomic analysis of the bacteria and the host. The analysis of dual RNA by RNA sequencing technology is challenging, due the difficulties of extracting bacterial RNA, which is overwhelmingly outnumbered by the host RNA, especially at the critical early time points post-infection (prior to 48 h). Here, we describe a novel technique that employed the infusion of an RNA preserving reagent (RNAlater) into the lungs of the animals, through the trachea, under deep anesthesia. This method enabled the isolation of stable dual mRNA from the lungs of mice infected with Y. pestis, as early as 24 h post-infection. The RNA was used for transcriptomic analysis, which provided a comprehensive gene expression profile of both the host and the pathogen. MDPI 2021-10-18 /pmc/articles/PMC8539884/ /pubmed/34683487 http://dx.doi.org/10.3390/microorganisms9102166 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Israeli, Ofir
Cohen-Gihon, Inbar
Aftalion, Moshe
Gur, David
Vagima, Yaron
Zauberman, Ayelet
Levy, Yinon
Zvi, Anat
Chitlaru, Theodor
Mamroud, Emanuelle
Tidhar, Avital
Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of Yersinia pestis Pulmonary Infection
title Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of Yersinia pestis Pulmonary Infection
title_full Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of Yersinia pestis Pulmonary Infection
title_fullStr Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of Yersinia pestis Pulmonary Infection
title_full_unstemmed Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of Yersinia pestis Pulmonary Infection
title_short Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of Yersinia pestis Pulmonary Infection
title_sort novel rna extraction method for dual rna-seq analysis of pathogen and host in the early stages of yersinia pestis pulmonary infection
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8539884/
https://www.ncbi.nlm.nih.gov/pubmed/34683487
http://dx.doi.org/10.3390/microorganisms9102166
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