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      Using click chemistry to study microbial ecology and evolution

      review-article
      1 , , 2 ,
      ISME Communications
      Nature Publishing Group UK
      Microbial ecology, Microbial ecology

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          Abstract

          Technological advances have largely driven the revolution in our understanding of the structure and function of microbial communities. Culturing, long the primary tool to probe microbial life, was supplanted by sequencing and other -omics approaches, which allowed detailed quantitative insights into species composition, metabolic potential, transcriptional activity, secretory responses and more. Although the ability to characterize “who’s there” has never been easier or cheaper, it remains technically challenging and expensive to understand what the diverse species and strains that comprise microbial communities are doing in situ, and how these behaviors change through time. Our aim in this brief review is to introduce a developing toolkit based on click chemistry that can accelerate and reduce the expense of functional analyses of the ecology and evolution of microbial communities. After first outlining the history of technological development in this field, we will discuss key applications to date using diverse labels, including BONCAT, and then end with a selective (biased) view of areas where click-chemistry and BONCAT-based approaches stand to have a significant impact on our understanding of microbial communities.

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          Most cited references85

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          Gut microbiota in human metabolic health and disease

          Observational findings achieved during the past two decades suggest that the intestinal microbiota may contribute to the metabolic health of the human host and, when aberrant, to the pathogenesis of various common metabolic disorders including obesity, type 2 diabetes, non-alcoholic liver disease, cardio-metabolic diseases and malnutrition. However, to gain a mechanistic understanding of how the gut microbiota affects host metabolism, research is moving from descriptive microbiota census analyses to cause-and-effect studies. Joint analyses of high-throughput human multi-omics data, including metagenomics and metabolomics data, together with measures of host physiology and mechanistic experiments in humans, animals and cells hold potential as initial steps in the identification of potential molecular mechanisms behind reported associations. In this Review, we discuss the current knowledge on how gut microbiota and derived microbial compounds may link to metabolism of the healthy host or to the pathogenesis of common metabolic diseases. We highlight examples of microbiota-targeted interventions aiming to optimize metabolic health, and we provide perspectives for future basic and translational investigations within the nascent and promising research field.
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            A chemical method for fast and sensitive detection of DNA synthesis in vivo.

            We have developed a method to detect DNA synthesis in proliferating cells, based on the incorporation of 5-ethynyl-2'-deoxyuridine (EdU) and its subsequent detection by a fluorescent azide through a Cu(I)-catalyzed [3 + 2] cycloaddition reaction ("click" chemistry). Detection of the EdU label is highly sensitive and can be accomplished in minutes. The small size of the fluorescent azides used for detection results in a high degree of specimen penetration, allowing the staining of whole-mount preparations of large tissue and organ explants. In contrast to BrdU, the method does not require sample fixation or DNA denaturation and permits good structural preservation. We demonstrate the use of the method in cultured cells and in the intestine and brain of whole animals.
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              Cell surface engineering by a modified Staudinger reaction.

              Selective chemical reactions enacted within a cellular environment can be powerful tools for elucidating biological processes or engineering novel interactions. A chemical transformation that permits the selective formation of covalent adducts among richly functionalized biopolymers within a cellular context is presented. A ligation modeled after the Staudinger reaction forms an amide bond by coupling of an azide and a specifically engineered triarylphosphine. Both reactive partners are abiotic and chemically orthogonal to native cellular components. Azides installed within cell surface glycoconjugates by metabolism of a synthetic azidosugar were reacted with a biotinylated triarylphosphine to produce stable cell-surface adducts. The tremendous selectivity of the transformation should permit its execution within a cell's interior, offering new possibilities for probing intracellular interactions.
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                Author and article information

                Contributors
                s.i.van.kasteren@chem.leidenuniv.nl
                d.e.rozen@biology.leidenuniv.nl
                Journal
                ISME Commun
                ISME Commun
                ISME Communications
                Nature Publishing Group UK (London )
                2730-6151
                2730-6151
                31 January 2023
                31 January 2023
                2023
                : 3
                : 9
                Affiliations
                [1 ]GRID grid.5132.5, ISNI 0000 0001 2312 1970, Leiden Institute of Chemistry and The Institute of Chemical Immunology, , Leiden University, ; Einsteinweg 55, Leiden, 2300 RA The Netherlands
                [2 ]GRID grid.5132.5, ISNI 0000 0001 2312 1970, Institute of Biology, , Leiden University, ; Sylviusweg 72, Leiden, 2300 RA The Netherlands
                Author information
                http://orcid.org/0000-0002-7772-0239
                Article
                205
                10.1038/s43705-022-00205-5
                9889756
                36721064
                012a0ebb-e292-4a92-bea5-7a24aa169919
                © The Author(s) 2023

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 29 June 2022
                : 16 November 2022
                : 23 November 2022
                Categories
                Review Article
                Custom metadata
                © The Author(s) 2023

                microbial ecology
                microbial ecology

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