{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T00:33:12Z","timestamp":1760315592013,"version":"build-2065373602"},"reference-count":19,"publisher":"Wiley","issue":"16","license":[{"start":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T00:00:00Z","timestamp":1690243200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemBioChem"],"published-print":{"date-parts":[[2023,8,15]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Polymicrobial infections involving various combinations of microorganisms, such as <jats:italic>Escherichia<\/jats:italic>, <jats:italic>Pseudomonas<\/jats:italic>, or <jats:italic>Yersinia<\/jats:italic>, can lead to acute and chronic diseases in for example the gastrointestinal and respiratory tracts. Our aim is to modulate microbial communities by targeting the posttranscriptional regulator system called carbon storage regulator A (CsrA) (or also repressor of secondary metabolites (RsmA)). In previous studies, we identified easily accessible CsrA binding scaffolds and macrocyclic CsrA binding peptides through biophysical screening and phage display technology. However, due to the lack of an appropriate <jats:italic>in bacterio<\/jats:italic> assay to evaluate the cellular effects of these inhibitor hits, the focus of the present study is to establish an <jats:italic>in bacterio<\/jats:italic> assay capable of probing and quantifying the impact on CsrA\u2010regulated cellular mechanisms. We have successfully developed an assay based on a luciferase reporter gene assay, which in combination with a qPCR expression gene assay, allows for the monitoring of expression levels of different downstream targets of CsrA. The chaperone protein CesT was used as a suitable positive control for the assay, and in time\u2010dependent experiments, we observed a CesT\u2010mediated increase in bioluminescence over time. By this means, the cellular on\u2010target effects of non\u2010bactericidal\/non\u2010bacteriostatic virulence modulating compounds targeting CsrA\/RsmA can be evaluated.<\/jats:p>","DOI":"10.1002\/cbic.202300369","type":"journal-article","created":{"date-parts":[[2023,7,12]],"date-time":"2023-07-12T06:13:51Z","timestamp":1689142431000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment of an In Bacterio Assay for the Assessment of Carbon Storage Regulator A (CsrA) Inhibitors"],"prefix":"10.1002","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4414-006X","authenticated-orcid":false,"given":"Yingwen","family":"Wu","sequence":"first","affiliation":[{"name":"Department of Antiviral &amp; Antivirulence Drugs (AVID) Helmholtz Institute for Pharmaceutical Research Saarland (HIPS) Saarland University  66123 Saarbr\u00fccken Germany"},{"name":"Cluster of Excellence RESIST (EXC 2155) Hanover Medical School  Carl-Neuberg-Stra\u00dfe 1 30625 Hanover Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9319-5999","authenticated-orcid":false,"given":"Ben G. E.","family":"Zoller","sequence":"additional","affiliation":[{"name":"Department of Antiviral &amp; Antivirulence Drugs (AVID) Helmholtz Institute for Pharmaceutical Research Saarland (HIPS) Saarland University  66123 Saarbr\u00fccken Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mohamed Ashraf Mostafa","family":"Kamal","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery (DDEL) Helmholtz Institute for Pharmaceutical Research Saarland (HIPS) Saarland University  66123 Saarbr\u00fccken Germany"},{"name":"Department of Pharmacy Saarland University  66123 Saarbr\u00fccken Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3394-0432","authenticated-orcid":false,"given":"Sven\u2010Kevin","family":"Hotop","sequence":"additional","affiliation":[{"name":"Department of Chemical Biology Helmholtz Centre for Infection Research German Center for Infection Research (DZIF)  38124 Braunschweig Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5864-8462","authenticated-orcid":false,"given":"Claus\u2010Michael","family":"Lehr","sequence":"additional","affiliation":[{"name":"Department of Drug Delivery (DDEL) Helmholtz Institute for Pharmaceutical Research Saarland (HIPS) Saarland University  66123 Saarbr\u00fccken Germany"},{"name":"Department of Pharmacy Saarland University  66123 Saarbr\u00fccken Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8971-7045","authenticated-orcid":false,"given":"Mark","family":"Br\u00f6nstrup","sequence":"additional","affiliation":[{"name":"Department of Chemical Biology Helmholtz Centre for Infection Research German Center for Infection Research (DZIF)  38124 Braunschweig Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8177-3280","authenticated-orcid":false,"given":"Petra","family":"Dersch","sequence":"additional","affiliation":[{"name":"Institute of Infectiology Center for Molecular Biology of Inflammation (ZMBE) University of M\u00fcnster  48149 M\u00fcnster Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0503-5830","authenticated-orcid":false,"given":"Martin","family":"Empting","sequence":"additional","affiliation":[{"name":"Department of Antiviral &amp; 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