{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T14:54:44Z","timestamp":1783004084730,"version":"3.54.5"},"reference-count":38,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,4,3]],"date-time":"2024-04-03T00:00:00Z","timestamp":1712102400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Syst. Biol."],"abstract":"<jats:p>Although blood sampling and medical imaging are well-established techniques in clinical diagnostics, they often require long post-processing procedures. Fast and simple quantification of signaling molecules can enable efficient health monitoring and improve diagnoses. Thyroid hormones (THs) treatment relies on trial-and-error dose adjustments, and requires constant tracking via blood tests. Thus, a fast and reliable method that can constantly track THs levels could substantially improve patient quality of life by reducing their visits to doctors. Synthetic biosensors have shown to be inexpensive and easy tools for sensing molecules, with their use in healthcare increasing over time. This study describes the construction of an engineered THs bacterial biosensor, consisting of a split-intein-based TH receptor ligand binding domain (LBD) biosensor that reconstitutes green fluorescence protein (GFP) after binding to TH. This biosensor could quantitatively measure THs concentrations by evaluating fluorescence intensity. <jats:italic>In vitro<\/jats:italic> sensing using <jats:italic>Escherichia coli<\/jats:italic> produced GFP over a wide dynamic range. The biosensor was further optimized by adding a double LBD, which enhanced its dynamic range until it reached healthy physiological conditions. Moreover, a mathematical model was developed to assess the dynamic properties of the biosensor and to provide a basis for the characterization of other intein-mediated biosensors. This type of biosensor can be used as the basis for novel treatments of thyroid diseases and can be adapted to measure the concentrations of other hormones, giving rise to a series of mathematically characterized modular biosensors.<\/jats:p>","DOI":"10.3389\/fsysb.2024.1270071","type":"journal-article","created":{"date-parts":[[2024,4,3]],"date-time":"2024-04-03T05:05:51Z","timestamp":1712120751000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["Intein-mediated thyroid hormone biosensors: towards controlled delivery of hormone therapy"],"prefix":"10.3389","volume":"4","author":[{"given":"Quim","family":"Mart\u00ed-Baena","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreu","family":"Pascuet-Fontanet","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomas","family":"Berjaga-Buisan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miriam","family":"Caravaca-Rodr\u00edguez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaume","family":"Puig-Costa-Juss\u00e0","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Avencia","family":"Sanchez-Mejias","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dimitrije","family":"Ivan\u010di\u0107","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sira","family":"Mogas-D\u00edez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc","family":"G\u00fcell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Javier","family":"Macia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,4,3]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"486","DOI":"10.1089\/1050725041517138","article-title":"Joint statement on the U.S. Food and Drug Administration's decision regarding bioequivalence of levothyroxine sodium","volume":"14","year":"2004","journal-title":"Thyroid"},{"key":"B2","doi-asserted-by":"publisher","first-page":"11005","DOI":"10.1073\/pnas.0904366106","article-title":"Modulation of intein activity by its neighboring extein substrates","volume":"106","author":"Amitai","year":"2009","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B3","doi-asserted-by":"publisher","first-page":"602","DOI":"10.3390\/s23020602","article-title":"A novel strategy for selective thyroid hormone determination based on an electrochemical biosensor with graphene nanocomposite","volume":"23","author":"Baluta","year":"2023","journal-title":"Sensors (Basel, Switz.)"},{"key":"B4","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1016\/j.mimet.2004.05.003","article-title":"An optical microsensor to measure fluorescent light intensity in biofilms","volume":"58","author":"Beyenal","year":"2004","journal-title":"J. Microbiol. Methods"},{"key":"B5","doi-asserted-by":"publisher","first-page":"1390","DOI":"10.1038\/s41467-021-21740-0","article-title":"Applications, challenges, and needs for employing synthetic biology beyond the lab","volume":"12","author":"Brooks","year":"2021","journal-title":"Nat. Commun."},{"key":"B6","volume-title":"Communication systems: an introduction to signals and noise in electrical communication","author":"Carlson","year":"2010"},{"key":"B7","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1007\/s12325-019-01080-8","article-title":"Hypothyroidism in context: where we\u2019ve been and where we\u2019re going","volume":"36","author":"Chiovato","year":"2019","journal-title":"Adv. Ther."},{"key":"B8","doi-asserted-by":"publisher","first-page":"361","DOI":"10.1038\/s41467-018-06034-2","article-title":"Valores de referencia de hormonas tiroideas, tirotropina y tiroglobulina en ni\u00f1os sanos zaragozanos","volume":"51","author":"Cort\u00e9s-Blanco","year":"1999","journal-title":"Esp Pediatr"},{"key":"B9","doi-asserted-by":"publisher","first-page":"63","DOI":"10.3389\/fmicb.2014.00063","article-title":"Fusion tags for protein solubility, purification, and immunogenicity in Escherichia coli: the novel fh8 system","volume":"5","author":"Costa","year":"2014","journal-title":"Front. Microbiol."},{"key":"B10","doi-asserted-by":"publisher","first-page":"100828","DOI":"10.1016\/j.nantod.2019.100828","article-title":"Skin in the diagnostics game: wearable biosensor nano- and microsystems for medical diagnostics","volume":"30","author":"Dervisevic","year":"2020","journal-title":"Nano Today"},{"key":"B11","volume-title":"Clinical methods: the history, physical, and laboratory examinations","author":"Dickson","year":"1990"},{"key":"B12","doi-asserted-by":"publisher","first-page":"351864","DOI":"10.1155\/2012\/351864","article-title":"Tsh and thyrotropic agonists: key actors in thyroid homeostasis","volume":"2012","author":"Dietrich","year":"2012","journal-title":"J. Thyroid Res."},{"key":"B13","doi-asserted-by":"publisher","first-page":"736","DOI":"10.1016\/j.ijmedinf.2010.08.006","article-title":"Effectiveness of telemedicine: a systematic review of reviews","volume":"79","author":"Ekeland","year":"2010","journal-title":"Int. J. Med. Inf."},{"key":"B14","doi-asserted-by":"publisher","first-page":"2052","DOI":"10.1210\/en.2017-00250","article-title":"Thyroid hormone signaling pathways: time for a more precise nomenclature","volume":"158","author":"Flamant","year":"2017","journal-title":"Endocrinology"},{"key":"B15","doi-asserted-by":"publisher","first-page":"238","DOI":"10.1159\/000493123","article-title":"European thyroid association (eta) and thyroid federation international (tfi) joint position statement on the interchangeability of levothyroxine products in eu countries","volume":"7","author":"Fliers","year":"2018","journal-title":"Eur. Thyroid J."},{"key":"B16","doi-asserted-by":"publisher","first-page":"2456","DOI":"10.1021\/ac020756b","article-title":"An in vitro screening system for protein splicing inhibitors based on green fluorescent protein as an indicator","volume":"75","author":"Gangopadhyay","year":"2003","journal-title":"Anal. Chem."},{"key":"B17","doi-asserted-by":"publisher","first-page":"247","DOI":"10.1016\/j.fob.2012.08.002","article-title":"Bacterial biosensors for screening isoform-selective ligands for human thyroid receptors \u03b1-1 and \u03b2-1","volume":"2","author":"Gierach","year":"2012","journal-title":"FEBS Open Bio"},{"key":"B18","doi-asserted-by":"publisher","first-page":"1328","DOI":"10.1021\/acssynbio.0c00010","article-title":"Two-component biosensors: unveiling the mechanisms of predictable tunability","volume":"9","author":"Gonzalez-Flo","year":"2020","journal-title":"ACS Synth. Biol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"48","DOI":"10.12659\/AJCR.883788","article-title":"Subcutaneous application of levothyroxine as successful treatment option in a patient with malabsorption","volume":"14","author":"Groener","year":"2013","journal-title":"Am. J. case Rep."},{"key":"B20","doi-asserted-by":"publisher","first-page":"916","DOI":"10.1016\/j.jmb.2005.09.088","article-title":"Minimization and stabilization of the Mycobacterium tuberculosis reca intein","volume":"354","author":"Hiraga","year":"2005","journal-title":"J. Mol. Biol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"544","DOI":"10.3389\/fendo.2019.00544","article-title":"Sustained release t3 therapy: animal models and translational applications","volume":"10","author":"Idrees","year":"2019","journal-title":"Front. Endocrinol."},{"key":"B22","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1038\/nrg2775","article-title":"Synthetic biology: applications come of age","volume":"11","author":"Khalil","year":"2010","journal-title":"Nat. Rev. Genet."},{"key":"B23","doi-asserted-by":"publisher","first-page":"36","DOI":"10.1016\/j.coelec.2020.03.003","article-title":"Trends and perspectives in dna biosensors as diagnostic devices","volume":"23","author":"Kowalczyk","year":"2020","journal-title":"Curr. Opin. Electrochem."},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4302-6014-1","article-title":"Sensor technologies: healthcare, wellness, and environmental applications","volume":"1","author":"McGrath","year":"2013","journal-title":"Sens. Technol. Healthc. Wellness Environ. Appl."},{"key":"B25","doi-asserted-by":"publisher","first-page":"124955","DOI":"10.1016\/j.ijbiomac.2023.124955","article-title":"Bacterial cellulose-based composites as vehicles for dermal and transdermal drug delivery: a review","volume":"242","author":"Mohammadi","year":"2023","journal-title":"Int. J. Biol. Macromol."},{"key":"B26","doi-asserted-by":"publisher","first-page":"5151","DOI":"10.1021\/ac000617z","article-title":"A fluorescent indicator for detecting protein-protein interactions in vivo based on protein splicing","volume":"72","author":"Ozawa","year":"2000","journal-title":"Anal. Chem."},{"key":"B27","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1038\/nbt1172","article-title":"Engineering and characterization of a superfolder green fluorescent protein","volume":"24","author":"P\u00e9delacq","year":"2006","journal-title":"Nat. Biotechnol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"1529","DOI":"10.1038\/s41467-020-15272-2","article-title":"An expanded library of orthogonal split inteins enables modular multi-peptide assemblies","volume":"11","author":"Pinto","year":"2020","journal-title":"Nat. Commun."},{"key":"B29","doi-asserted-by":"publisher","first-page":"68","DOI":"10.1016\/j.coi.2016.06.004","article-title":"Chimeric antigen receptors: driving immunology towards synthetic biology","volume":"41","author":"Sadelain","year":"2016","journal-title":"Curr. Opin. Immunol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"446","DOI":"10.1039\/C3SC52951G","article-title":"Inteins: nature\u2019s gift to protein chemists","volume":"5","author":"Shah","year":"2014","journal-title":"Chem. Sci."},{"key":"B31","doi-asserted-by":"publisher","first-page":"8538","DOI":"10.1073\/pnas.1701083114","article-title":"A promiscuous split intein with expanded protein engineering applications","volume":"114","author":"Stevens","year":"2017","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B32","doi-asserted-by":"publisher","first-page":"121011","DOI":"10.1016\/j.ijpharm.2021.121011","article-title":"Poly(caprolactone)-based subcutaneous implant for sustained delivery of levothyroxine","volume":"607","author":"Stewart","year":"2021","journal-title":"Int. J. Pharm."},{"key":"B33","doi-asserted-by":"publisher","first-page":"64","DOI":"10.1186\/s12915-019-0685-x","article-title":"Synthetic biology goes cell-free","volume":"17","author":"Tinafar","year":"2019","journal-title":"BMC Biol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"eaav4523","DOI":"10.1126\/scitranslmed.aav4523","article-title":"Intein-mediated protein trans-splicing expands adeno-associated virus transfer capacity in the retina","volume":"11","author":"Tornabene","year":"2019","journal-title":"Sci. Transl. Med."},{"key":"B35","doi-asserted-by":"publisher","first-page":"810180","DOI":"10.3389\/fbioe.2022.810180","article-title":"Protein splicing of inteins: a powerful tool in synthetic biology","volume":"10","author":"Wang","year":"2022","journal-title":"Front. Bioeng. Biotechnol."},{"key":"B36","doi-asserted-by":"publisher","first-page":"3491","DOI":"10.1210\/jc.2018-02794","article-title":"Insight into molecular determinants of t3 vs t4 recognition from mutations in thyroid hormone receptor \u03b1 and \u03b2","volume":"104","author":"Wejaphikul","year":"2019","journal-title":"J. Clin. Endocrinol. Metabolism"},{"key":"B37","doi-asserted-by":"publisher","first-page":"889","DOI":"10.1038\/12879","article-title":"A genetic system yields self-cleaving inteins for bioseparations","volume":"17","author":"Wood","year":"1999","journal-title":"Nat. Biotechnol."},{"key":"B38","doi-asserted-by":"publisher","first-page":"1097","DOI":"10.1152\/physrev.2001.81.3.1097","article-title":"Physiological and molecular basis of thyroid hormone action","volume":"81","author":"Yen","year":"2001","journal-title":"Physiol. Rev."}],"container-title":["Frontiers in Systems Biology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fsysb.2024.1270071\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,4,3]],"date-time":"2024-04-03T05:05:54Z","timestamp":1712120754000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fsysb.2024.1270071\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,4,3]]},"references-count":38,"alternative-id":["10.3389\/fsysb.2024.1270071"],"URL":"https:\/\/doi.org\/10.3389\/fsysb.2024.1270071","relation":{},"ISSN":["2674-0702"],"issn-type":[{"value":"2674-0702","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,4,3]]},"article-number":"1270071"}}