Co-transcriptomic Analysis by RNA Sequencing to Simultaneously Measure Regulated Gene Expression in Host and Bacterial Pathogen

Timothy Ravasi, Charalampos Harris Mavromatis, Nilesh J. Bokil, Mark A. Schembri, Matthew J. Sweet

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Intramacrophage pathogens subvert antimicrobial defence pathways using various mechanisms, including the targeting of host TLR-mediated transcriptional responses. Conversely, TLR-inducible host defence mechanisms subject intramacrophage pathogens to stress, thus altering pathogen gene expression programs. Important biological insights can thus be gained through the analysis of gene expression changes in both the host and the pathogen during an infection. Traditionally, research methods have involved the use of qPCR, microarrays and/or RNA sequencing to identify transcriptional changes in either the host or the pathogen. Here we describe the application of RNA sequencing using samples obtained from in vitro infection assays to simultaneously quantify both host and bacterial pathogen gene expression changes, as well as general approaches that can be undertaken to interpret the RNA sequencing data that is generated. These methods can be used to provide insights into host TLR-regulated transcriptional responses to microbial challenge, as well as pathogen subversion mechanisms against such responses.
Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherSpringer Nature
Pages145-158
Number of pages14
ISBN (Print)978-1-4939-3333-4
DOIs
StatePublished - Jan 24 2016

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

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