Dual RNA-seq analysis of in vitro infection multiplicity and RNA depletion methods in Chlamydia-infected epithelial cells
Date
2021-05-17Journal
Scientific ReportsPublisher
Springer NatureType
Article
Metadata
Show full item recordAbstract
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 Chlamydia trachomatis infected epithelial cells to examine transcriptomic responses from both organisms. We compared two time points post infection (1 and 24 h), 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. © 2021, The Author(s).Keyword
dual RNA-seqChlamydia trachomatis--genetics
Host-Pathogen Interactions--genetics
Sequence Analysis, RNA
Identifier to cite or link to this item
http://hdl.handle.net/10713/15881ae974a485f413a2113503eed53cd6c53
10.1038/s41598-021-89921-x
Scopus Count
Collections
Related articles
- Early Transcriptional Landscapes of <i>Chlamydia trachomatis</i>-Infected Epithelial Cells at Single Cell Resolution.
- Authors: Hayward RJ, Marsh JW, Humphrys MS, Huston WM, Myers GSA
- Issue date: 2019
- Dual RNA-Seq of Chlamydia and Host Cells.
- Authors: Marsh JW, Hayward RJ, Shetty A, Mahurkar A, Humphrys MS, Myers GSA
- Issue date: 2019
- Bioinformatic analysis of bacteria and host cell dual RNA-sequencing experiments.
- Authors: Marsh JW, Hayward RJ, Shetty AC, Mahurkar A, Humphrys MS, Myers GSA
- Issue date: 2018 Nov 27
- High dynamic range detection of Chlamydia trachomatis growth by direct quantitative PCR of the infected cells.
- Authors: Eszik I, Lantos I, Önder K, Somogyvári F, Burián K, Endrész V, Virok DP
- Issue date: 2016 Jan
- Host cell response and distinct gene expression profiles at different stages of Chlamydia trachomatis infection reveals stage-specific biomarkers of infection.
- Authors: Dzakah EE, Huang L, Xue Y, Wei S, Wang X, Chen H, Shui J, Kyei F, Rashid F, Zheng H, Yang B, Tang S
- Issue date: 2021 Jan 4