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      Exploring the transcriptional landscape of phage–host interactions using novel high-throughput approaches

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          Abstract

          In the last decade, powerful high-throughput sequencing approaches have emerged to analyse microbial transcriptomes at a global scale. However, to date, applications of these approaches to microbial viruses such as phages remain scarce. Tailoring these techniques to virus-infected bacteria promises to obtain a detailed picture of the underexplored RNA biology and molecular processes during infection. In addition, transcriptome study of stress and perturbations induced by phages in their infected bacterial hosts is likely to reveal new fundamental mechanisms of bacterial metabolism and gene regulation. Here, we provide references and blueprints to implement emerging transcriptomic approaches towards addressing transcriptome architecture, RNA–RNA and RNA–protein interactions, RNA modifications, structures and heterogeneity of transcription profiles in infected cells that will provide guides for future directions in phage-centric therapeutic applications and microbial synthetic biology.

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          Nanopore sequencing technology, bioinformatics and applications

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            Phage diversity, genomics and phylogeny

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              Term-seq reveals abundant ribo-regulation of antibiotics resistance in bacteria.

              Riboswitches and attenuators are cis-regulatory RNA elements, most of which control bacterial gene expression via metabolite-mediated, premature transcription termination. We developed an unbiased experimental approach for genome-wide discovery of such ribo-regulators in bacteria. We also devised an experimental platform that quantitatively measures the in vivo activity of all such regulators in parallel and enables rapid screening for ribo-regulators that respond to metabolites of choice. Using this approach, we detected numerous antibiotic-responsive ribo-regulators that control antibiotic resistance genes in pathogens and in the human microbiome. Studying one such regulator in Listeria monocytogenes revealed an attenuation mechanism mediated by antibiotic-stalled ribosomes. Our results expose broad roles for conditional termination in regulating antibiotic resistance and provide a tool for discovering riboswitches and attenuators that respond to previously unknown ligands.
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                Author and article information

                Contributors
                Journal
                Curr Opin Microbiol
                Curr Opin Microbiol
                Current Opinion in Microbiology
                Current Biology
                1369-5274
                1879-0364
                1 February 2024
                February 2024
                : 77
                : 102419
                Affiliations
                [1 ]Department of Biosystems, Laboratory of Gene Technology, KU Leuven, Leuven, Belgium
                [2 ]Institute for Molecular Infection Biology (IMIB), Medical Faculty, University of Würzburg, Würzburg, Germany
                [3 ]Centre of Biological Engineering, University of Minho, Braga, Portugal
                [4 ]Helmholtz Institute for RNA-based Infection Research (HIRI), Helmholtz Centre for Infection Research (HZI), Würzburg, Germany
                Author notes
                [*]

                These authors contributed equally.

                Article
                S1369-5274(23)00156-X 102419
                10.1016/j.mib.2023.102419
                10884466
                38271748
                3e49b589-f0d4-4e6f-8d56-866461f98ddf
                © 2024 The Authors

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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                Microbiology & Virology
                Microbiology & Virology

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