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

被引:1
|
作者
Israeli, Ofir [1 ]
Cohen-Gihon, Inbar [1 ]
Aftalion, Moshe [1 ]
Gur, David [1 ]
Vagima, Yaron [1 ]
Zauberman, Ayelet [1 ]
Levy, Yinon [1 ]
Zvi, Anat [1 ]
Chitlaru, Theodor [1 ]
Mamroud, Emanuelle [1 ]
Tidhar, Avital [1 ]
机构
[1] Israel Inst Biol Res, Dept Biochem & Mol Genet, IL-74100 Ness Ziona, Israel
关键词
dual RNA-seq; pneumonic plague; Y; pestis; mouse model; infection;
D O I
10.3390/microorganisms9102166
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页数:8
相关论文
共 50 条
  • [1] Dual RNA-seq of pathogen and host
    Westermann, Alexander J.
    Gorski, Stanislaw A.
    Vogel, Joerg
    NATURE REVIEWS MICROBIOLOGY, 2012, 10 (09) : 618 - 630
  • [2] Dual RNA-seq of pathogen and host
    Alexander J. Westermann
    Stanislaw A. Gorski
    Jörg Vogel
    Nature Reviews Microbiology, 2012, 10 : 618 - 630
  • [3] Dual RNA-seq for host–pathogen transcriptomics
    Ross Cloney
    Nature Reviews Genetics, 2016, 17 : 126 - 127
  • [4] RNA CoMPASS: A Dual Approach for Pathogen and Host Transcriptome Analysis of RNA-Seq Datasets
    Xu, Guorong
    Strong, Michael J.
    Lacey, Michelle R.
    Baribault, Carl
    Flemington, Erik K.
    Taylor, Christopher M.
    PLOS ONE, 2014, 9 (02):
  • [5] Dual RNA-seq unveils noncoding RNA functions in host–pathogen interactions
    Alexander J. Westermann
    Konrad U. Förstner
    Fabian Amman
    Lars Barquist
    Yanjie Chao
    Leon N. Schulte
    Lydia Müller
    Richard Reinhardt
    Peter F. Stadler
    Jörg Vogel
    Nature, 2016, 529 : 496 - 501
  • [6] Host–pathogen duels revealed by dual RNA-seq in vivo
    Darren J. Burgess
    Nature Reviews Genetics, 2017, 18 : 143 - 143
  • [7] Resolving host-pathogen interactions by dual RNA-seq
    Westermann, Alexander J.
    Barquist, Lars
    Vogel, Joerg
    PLOS PATHOGENS, 2017, 13 (02)
  • [8] RNA-Seq Analysis Illuminates the Early Stages of Plasmodium Liver Infection
    Toro-Moreno, Maria
    Sylvester, Kayla
    Srivastava, Tamanna
    Posfai, Dora
    Derbyshire, Emily R.
    MBIO, 2020, 11 (01):
  • [9] Determination of sRNA Expressions by RNA-seq in Yersinia pestis Grown In Vitro and during Infection
    Yan, Yanfeng
    Su, Shanchun
    Meng, Xiangrong
    Ji, Xiaolan
    Qu, Yi
    Liu, Zizhong
    Wang, Xiaoyi
    Cui, Yujun
    Deng, Zhongliang
    Zhou, Dongsheng
    Jiang, Wencan
    Yang, Ruifu
    Han, Yanping
    PLOS ONE, 2013, 8 (09):
  • [10] Dual RNA-seq unveils noncoding RNA functions in host-pathogen interactions
    Westermann, Alexander J.
    Foerstner, Konrad U.
    Amman, Fabian
    Barquist, Lars
    Chao, Yanjie
    Schulte, Leon N.
    Mueller, Lydia
    Reinhardt, Richard
    Stadler, Peter F.
    Vogel, Joerg
    NATURE, 2016, 529 (7587) : 496 - +