Dual RNA-Seq analysis of in vitro infection multiplicity in Chlamydia-infected epithelial cells

Hayward, RJ; Humphrys, MS; Huston, WM; Myers, GSA

Dual RNA-Seq analysis of in vitro infection multiplicity in Chlamydia-infected epithelial cells

Hayward, RJ Humphrys, MS Huston, WM Myers, GSA

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Publisher:: Cold Spring Harbor Laboratory
Publication Type:: Journal Article

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Field	Value	Language
dc.contributor.author	Hayward, RJ
dc.contributor.author	Humphrys, MS
dc.contributor.author	Huston, WM
dc.contributor.author	Myers, GSA
dc.date.accessioned	2020-11-10T04:09:16Z
dc.date.available	2020-11-10T04:09:16Z
dc.identifier.uri	http://hdl.handle.net/10453/143872
dc.description.abstract	<jats:title>Abstract</jats:title><jats:p>Dual RNA-seq experiments examining viral and bacterial pathogens are increasing, but vary considerably in their experimental designs, such as infection rates and RNA depletion methods. Here, we have applied dual RNA-seq to <jats:italic>Chlamydia trachomatis</jats:italic> infected epithelial cells to examine transcriptomic responses from both organisms. We compared two time points post infection (1 and 24 hours), three multiplicity of infection (MOI) ratios (0.1, 1 and 10) and two RNA depletion methods (rRNA and polyA). Capture of bacterial-specific RNA were greatest when combining rRNA and polyA depletion, and when using a higher MOI. However, under these conditions, host RNA capture was negatively impacted. Although it is tempting to use high infection rates, the implications on host cell survival, the potential reduced length of infection cycles and real world applicability should be considered. This data highlights the delicate nature of balancing host-pathogen RNA capture and will assist future transcriptomic-based studies to achieve more specific and relevant infection-related biological insights.</jats:p>
dc.language	en
dc.publisher	Cold Spring Harbor Laboratory
dc.relation.isbasedon	10.1101/2020.10.21.347906
dc.rights	info:eu-repo/semantics/openAccess
dc.title	Dual RNA-Seq analysis of in vitro infection multiplicity in Chlamydia-infected epithelial cells
dc.type	Journal Article
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	open_access	*
dc.date.updated	2020-11-10T04:09:06Z

Abstract:

AbstractDual RNA-seq experiments examining viral and bacterial pathogens are increasing, but vary considerably in their experimental designs, such as infection rates and RNA depletion methods. Here, we have applied dual RNA-seq to Chlamydia trachomatis infected epithelial cells to examine transcriptomic responses from both organisms. We compared two time points post infection (1 and 24 hours), three multiplicity of infection (MOI) ratios (0.1, 1 and 10) and two RNA depletion methods (rRNA and polyA). Capture of bacterial-specific RNA were greatest when combining rRNA and polyA depletion, and when using a higher MOI. However, under these conditions, host RNA capture was negatively impacted. Although it is tempting to use high infection rates, the implications on host cell survival, the potential reduced length of infection cycles and real world applicability should be considered. This data highlights the delicate nature of balancing host-pathogen RNA capture and will assist future transcriptomic-based studies to achieve more specific and relevant infection-related biological insights.

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http://hdl.handle.net/10453/143872