{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T05:45:59Z","timestamp":1773294359558,"version":"3.50.1"},"reference-count":42,"publisher":"Oxford University Press (OUP)","issue":"15","license":[{"start":{"date-parts":[[2023,7,27]],"date-time":"2023-07-27T00:00:00Z","timestamp":1690416000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010663","name":"European Research Council","doi-asserted-by":"publisher","award":["785800"],"award-info":[{"award-number":["785800"]}],"id":[{"id":"10.13039\/100010663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001711","name":"Swiss National Science Foundation","doi-asserted-by":"publisher","award":["59511.1 IP-LS"],"award-info":[{"award-number":["59511.1 IP-LS"]}],"id":[{"id":"10.13039\/501100001711","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003006","name":"ETH Zurich","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003006","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,8,25]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Synthetic biology holds great promise to improve the safety and efficacy of future gene and engineered cell therapies by providing new means of endogenous or exogenous control of the embedded therapeutic programs. Here, we focused on gluconate as a clinically licensed small-molecule inducer and engineered gluconate-sensitive molecular switches to regulate transgene expression in human cell cultures and in mice. Several switch designs were assembled based on the gluconate-responsive transcriptional repressor GntR from Escherichia coli. Initially we assembled OFF- and ON-type switches by rewiring the native gluconate-dependent binding of GntR to target DNA sequences in mammalian cells. Then, we utilized the ability of GntR to dimerize in the presence of gluconate to activate gene expression from a split transcriptional activator. By means of random mutagenesis of GntR combined with phenotypic screening, we identified variants that significantly enhanced the functionality of the genetic devices, enabling the construction of robust two-input logic gates. We also demonstrated the potential utility of the synthetic switch in two in vivo settings, one employing implantation of alginate-encapsulated engineered cells and the other involving modification of host cells by DNA delivery. Then, as proof-of-concept, the gluconate-actuated genetic switch was connected to insulin secretion, and the components encoding gluconate-induced insulin production were introduced into\u00a0type-1 diabetic mice\u00a0as naked DNA via hydrodynamic tail vein injection. Normoglycemia was restored, thereby showcasing the suitability of oral gluconate to regulate in situ production of a therapeutic protein.<\/jats:p>","DOI":"10.1093\/nar\/gkad600","type":"journal-article","created":{"date-parts":[[2023,7,27]],"date-time":"2023-07-27T10:48:09Z","timestamp":1690454889000},"page":"e85-e85","source":"Crossref","is-referenced-by-count":12,"title":["Evolution of molecular switches for regulation of transgene expression by clinically licensed gluconate"],"prefix":"10.1093","volume":"51","author":[{"given":"Ana Palma","family":"Teixeira","sequence":"first","affiliation":[{"name":"Department of Biosystems Science and Engineering, ETH Zurich , Mattenstrasse 26 , CH-4058 Basel , Switzerland"}]},{"given":"Shuai","family":"Xue","sequence":"additional","affiliation":[{"name":"Department of Biosystems Science and Engineering, ETH Zurich , Mattenstrasse 26 , CH-4058 Basel , Switzerland"}]},{"given":"Jinbo","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Biosystems Science and Engineering, ETH Zurich , Mattenstrasse 26 , CH-4058 Basel , Switzerland"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8545-667X","authenticated-orcid":false,"given":"Martin","family":"Fussenegger","sequence":"additional","affiliation":[{"name":"Department of Biosystems Science and Engineering, ETH Zurich , Mattenstrasse 26 , CH-4058 Basel , Switzerland"},{"name":"Faculty of Science, University of Basel , Mattenstrasse 26 , CH-4058 Basel , Switzerland"}]}],"member":"286","published-online":{"date-parts":[[2023,7,27]]},"reference":[{"key":"2023082505041397600_B1","doi-asserted-by":"crossref","first-page":"1602","DOI":"10.1016\/j.drudis.2021.03.021","article-title":"The era of gene therapy: from preclinical development to clinical application","volume":"26","author":"Alhakamy","year":"2021","journal-title":"Drug Discov. 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Biotechnol."},{"key":"2023082505041397600_B8","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1158\/2326-6066.CIR-16-0043","article-title":"A Tet-On inducible system for controlling CD19-chimeric antigen receptor expression upon drug administration","volume":"4","author":"Sakemura","year":"2016","journal-title":"Cancer Immunol. Res."},{"key":"2023082505041397600_B9","doi-asserted-by":"crossref","first-page":"6295","DOI":"10.1126\/scitranslmed.abb6295","article-title":"Reversible ON-and OFF-switch chimeric antigen receptors controlled by lenalidomide","volume":"13","author":"Jan","year":"2021","journal-title":"Sci. Transl. 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Gene Med."},{"key":"2023082505041397600_B35","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1126\/science.aab2956","article-title":"Dynamics of epigenetic regulation at the single-cell level","volume":"351","author":"Bintu","year":"2016","journal-title":"Science"},{"key":"2023082505041397600_B36","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1634\/theoncologist.12-2-231","article-title":"Intravenous ferric gluconate significantly improves response to epoetin alfa versus oral iron or no iron in anemic patients with cancer receiving chemotherapy","volume":"12","author":"Henry","year":"2007","journal-title":"Oncologist"},{"key":"2023082505041397600_B37","doi-asserted-by":"crossref","first-page":"2020","DOI":"10.1016\/j.cell.2020.11.024","article-title":"High-throughput discovery and characterization of human transcriptional effectors","volume":"183","author":"Tycko","year":"2020","journal-title":"Cell"},{"key":"2023082505041397600_B38","doi-asserted-by":"crossref","first-page":"1891","DOI":"10.1016\/j.procbio.2016.08.028","article-title":"Gluconic acid: properties, production methods and applications\u2014An excellent opportunity for agro-industrial by-products and waste bio-valorization","volume":"51","author":"Canete-Rodriguez","year":"2016","journal-title":"Process Biochem."},{"key":"2023082505041397600_B39","doi-asserted-by":"crossref","first-page":"572","DOI":"10.1016\/j.omtm.2021.01.002","article-title":"iMATCH: an integrated modular assembly system for therapeutic combination high-capacity adenovirus gene therapy","volume":"20","author":"Br\u00fccher","year":"2021","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"2023082505041397600_B40","doi-asserted-by":"crossref","first-page":"6110","DOI":"10.1038\/s41467-022-33787-8","article-title":"Ex vivo engineered human plasma cells exhibit robust protein secretion and long-term engraftment in vivo","volume":"13","author":"Cheng","year":"2022","journal-title":"Nat. Commun."},{"key":"2023082505041397600_B41","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.tips.2020.11.008","article-title":"Biological cells as therapeutic delivery vehicles","volume":"42","author":"Bush","year":"2021","journal-title":"Trends Pharmacol. 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