Marsh James W, Hayward Regan J, Shetty Amol, Mahurkar Anup, Humphrys Michael S, Myers Garry S A
The iThree Institute, University of Technology Sydney, Ultimo, NSW, Australia.
Department of Microbiome Science, Max Planck Institute for Developmental Biology, Tübingen, Germany.
Methods Mol Biol. 2019;2042:123-135. doi: 10.1007/978-1-4939-9694-0_9.
During the infection of a host cell by a bacterial pathogen, a cascading series of gene expression changes occurs as each organism manipulates or responds to the other via defense or survival strategies. Unraveling this complex interplay is key for our understanding of bacterial virulence and host response pathways for the development of novel therapeutics. Dual RNA sequencing (dual RNA-Seq) has recently been developed to simultaneously capture host and bacterial transcriptomes from an infected cell. Leveraging the sensitivity and resolution allowed by RNA-seq, dual RNA-Seq can be applied to any bacteria-eukaryotic host interaction. We pioneered dual RNA-Seq to simultaneously capture Chlamydia and host expression profiles during an in vitro infection as proof of principle. Here we provide a detailed laboratory protocol and bioinformatics analysis guidelines for dual RNA-seq experiments focusing on Chlamydia as the organism of interest.
在细菌病原体感染宿主细胞的过程中，随着每种生物体通过防御或生存策略相互操纵或做出反应，会发生一系列级联的基因表达变化。揭示这种复杂的相互作用对于我们理解细菌毒力和宿主反应途径以开发新型治疗方法至关重要。双RNA测序（dual RNA-Seq）最近已被开发出来，用于同时从受感染细胞中捕获宿主和细菌转录组。利用RNA测序所允许的灵敏度和分辨率，双RNA测序可应用于任何细菌与真核宿主的相互作用。我们率先开展双RNA测序，以在体外感染期间同时捕获衣原体和宿主的表达谱，作为原理验证。在此，我们提供了一份详细的实验室方案和生物信息学分析指南，用于以衣原体为研究对象的双RNA测序实验。
