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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

Authors :
Ofir Israeli
Inbar Cohen-Gihon
Moshe Aftalion
David Gur
Yaron Vagima
Ayelet Zauberman
Yinon Levy
Anat Zvi
Theodor Chitlaru
Emanuelle Mamroud
Avital Tidhar
Source :
Microorganisms, Vol 9, Iss 10, p 2166 (2021)
Publication Year :
2021
Publisher :
MDPI AG, 2021.

Abstract

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.

Details

Language :
English
ISSN :
20762607
Volume :
9
Issue :
10
Database :
Directory of Open Access Journals
Journal :
Microorganisms
Publication Type :
Academic Journal
Accession number :
edsdoj.797869f328d426e8f385e653ac08a35
Document Type :
article
Full Text :
https://doi.org/10.3390/microorganisms9102166