Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of <i>Yersinia pestis</i> Pulmonary Infection
Pneumonic plague, caused by <i>Yersinia pestis,</i> 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...
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MDPI AG
2021-10-01
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author | Ofir Israeli Inbar Cohen-Gihon Moshe Aftalion David Gur Yaron Vagima Ayelet Zauberman Yinon Levy Anat Zvi Theodor Chitlaru Emanuelle Mamroud Avital Tidhar |
author_facet | Ofir Israeli Inbar Cohen-Gihon Moshe Aftalion David Gur Yaron Vagima Ayelet Zauberman Yinon Levy Anat Zvi Theodor Chitlaru Emanuelle Mamroud Avital Tidhar |
author_sort | Ofir Israeli |
collection | DOAJ |
description | Pneumonic plague, caused by <i>Yersinia pestis,</i> 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 <i>Y. pestis</i>, 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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issn | 2076-2607 |
language | English |
last_indexed | 2024-03-10T06:22:18Z |
publishDate | 2021-10-01 |
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series | Microorganisms |
spelling | doaj.art-a797869f328d426e8f385e653ac08a352023-11-22T19:15:25ZengMDPI AGMicroorganisms2076-26072021-10-01910216610.3390/microorganisms9102166Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of <i>Yersinia pestis</i> Pulmonary InfectionOfir Israeli0Inbar Cohen-Gihon1Moshe Aftalion2David Gur3Yaron Vagima4Ayelet Zauberman5Yinon Levy6Anat Zvi7Theodor Chitlaru8Emanuelle Mamroud9Avital Tidhar10Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelDepartment of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelDepartment of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelDepartment of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelDepartment of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelDepartment of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelDepartment of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelDepartment of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelDepartment of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelDepartment of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelDepartment of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelPneumonic plague, caused by <i>Yersinia pestis,</i> 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 <i>Y. pestis</i>, 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.https://www.mdpi.com/2076-2607/9/10/2166dual RNA-seqpneumonic plague<i>Y. pestis</i>mouse modelinfection |
spellingShingle | Ofir Israeli Inbar Cohen-Gihon Moshe Aftalion David Gur Yaron Vagima Ayelet Zauberman Yinon Levy Anat Zvi Theodor Chitlaru Emanuelle Mamroud Avital Tidhar Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of <i>Yersinia pestis</i> Pulmonary Infection Microorganisms dual RNA-seq pneumonic plague <i>Y. pestis</i> mouse model infection |
title | Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of <i>Yersinia pestis</i> Pulmonary Infection |
title_full | Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of <i>Yersinia pestis</i> Pulmonary Infection |
title_fullStr | Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of <i>Yersinia pestis</i> Pulmonary Infection |
title_full_unstemmed | Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of <i>Yersinia pestis</i> Pulmonary Infection |
title_short | Novel RNA Extraction Method for Dual RNA-seq Analysis of Pathogen and Host in the Early Stages of <i>Yersinia pestis</i> Pulmonary Infection |
title_sort | novel rna extraction method for dual rna seq analysis of pathogen and host in the early stages of i yersinia pestis i pulmonary infection |
topic | dual RNA-seq pneumonic plague <i>Y. pestis</i> mouse model infection |
url | https://www.mdpi.com/2076-2607/9/10/2166 |
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