A laboratory methodology for dual RNA-sequencing of bacteria and their host cells in vitro

Marsh, JW; Humphrys, MS; Myers, GSA

A laboratory methodology for dual RNA-sequencing of bacteria and their host cells in vitro

Marsh, JW

Humphrys, MS Myers, GSA

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Publication Type:: Journal Article
Citation:: Frontiers in Microbiology, 2017, 8 (SEP)
Issue Date:: 2017-09-21

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Field	Value	Language
dc.contributor.author	Marsh, JW https://orcid.org/0000-0003-4681-1012	en_US
dc.contributor.author	Humphrys, MS	en_US
dc.contributor.author	Myers, GSA https://orcid.org/0000-0002-4756-4810	en_US
dc.date.available	2017-09-06	en_US
dc.date.issued	2017-09-21	en_US
dc.identifier.citation	Frontiers in Microbiology, 2017, 8 (SEP)	en_US
dc.identifier.uri	http://hdl.handle.net/10453/117501
dc.description.abstract	© 2017 Marsh, Humphrys and Myers. Dual RNA-Sequencing leverages established next-generation sequencing (NGS)-enabled RNA-Seq approaches to measure genome-wide transcriptional changes of both an infecting bacteria and host cells. By simultaneously investigating both organisms from the same biological sample, dual RNA-Seq can provide unique insight into bacterial infection processes and reciprocal host responses at once. However, the difficulties involved in handling both prokaryotic and eukaryotic material require distinct, optimized procedures. We previously developed and applied dual RNA-Seq to measure prokaryotic and eukaryotic expression profiles of human cells infected with bacteria, using in vitro Chlamydia-infected epithelial cells as proof of principle. Here we provide a detailed laboratory protocol for in vitro dual RNA-Seq that is readily adaptable to any host-bacteria system of interest.	en_US
dc.relation.ispartof	Frontiers in Microbiology	en_US
dc.relation.isbasedon	10.3389/fmicb.2017.01830	en_US
dc.title	A laboratory methodology for dual RNA-sequencing of bacteria and their host cells in vitro	en_US
dc.type	Journal Article
utslib.citation.volume	SEP	en_US
utslib.citation.volume	8	en_US
utslib.for	0502 Environmental Science and Management	en_US
utslib.for	0503 Soil Sciences	en_US
utslib.for	0605 Microbiology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	open_access
pubs.issue	SEP	en_US
pubs.publication-status	Published	en_US
pubs.volume	8	en_US

Abstract:

© 2017 Marsh, Humphrys and Myers. Dual RNA-Sequencing leverages established next-generation sequencing (NGS)-enabled RNA-Seq approaches to measure genome-wide transcriptional changes of both an infecting bacteria and host cells. By simultaneously investigating both organisms from the same biological sample, dual RNA-Seq can provide unique insight into bacterial infection processes and reciprocal host responses at once. However, the difficulties involved in handling both prokaryotic and eukaryotic material require distinct, optimized procedures. We previously developed and applied dual RNA-Seq to measure prokaryotic and eukaryotic expression profiles of human cells infected with bacteria, using in vitro Chlamydia-infected epithelial cells as proof of principle. Here we provide a detailed laboratory protocol for in vitro dual RNA-Seq that is readily adaptable to any host-bacteria system of interest.

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http://hdl.handle.net/10453/117501