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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...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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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. |
format | Online Article Text |
id | pubmed-8539884 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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