{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T05:24:41Z","timestamp":1768800281980,"version":"3.49.0"},"reference-count":68,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2024,3,21]],"date-time":"2024-03-21T00:00:00Z","timestamp":1710979200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004040","name":"KU Leuven","doi-asserted-by":"publisher","award":["C14\/17\/067"],"award-info":[{"award-number":["C14\/17\/067"]}],"id":[{"id":"10.13039\/501100004040","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2024,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Estrogen receptor \u03b1 is commonly used in synthetic biology to control the activity of genome editing tools. The activating ligands, estrogens, however, interfere with various cellular processes, thereby limiting the applicability of this receptor. Altering its ligand preference to chemicals of choice solves this hurdle but requires adaptation of unspecified ligand\u2010interacting residues. Here, we provide a solution by combining rational protein design with multi\u2010site\u2010directed mutagenesis and directed evolution of stably integrated variants in <jats:italic>Saccharomyces cerevisiae.<\/jats:italic> This method yielded an estrogen receptor variant, named TERRA, that lost its estrogen responsiveness and became activated by tamoxifen, an anti\u2010estrogenic drug used for breast cancer treatment. This tamoxifen preference of TERRA was maintained in mammalian cells and mice, even when fused to Cre recombinase, expanding the mammalian synthetic biology toolbox. Not only is our platform transferable to engineer ligand preference of any steroid receptor, it can also profile drug\u2010resistance landscapes for steroid receptor\u2010targeted therapies.<\/jats:p>","DOI":"10.1002\/pro.4940","type":"journal-article","created":{"date-parts":[[2024,3,21]],"date-time":"2024-03-21T05:55:16Z","timestamp":1711000516000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Rational evolution for altering the ligand preference of estrogen receptor alpha"],"prefix":"10.1002","volume":"33","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9661-8312","authenticated-orcid":false,"given":"Roy","family":"Eerlings","sequence":"first","affiliation":[{"name":"Molecular Endocrinology Laboratory, Department of Cellular and Molecular Medicine KU Leuven  Leuven Belgium"},{"name":"Laboratory of Systems Biology VIB\u2010KU Leuven Center for Microbiology  Leuven Belgium"},{"name":"Laboratory for Genetics and Genomics, Center of Microbial and Plant Genetics, Department M2S KU Leuven  Heverlee Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Purvi","family":"Gupta","sequence":"additional","affiliation":[{"name":"Laboratory of Biomolecular Modelling and Design, Department of Chemistry KU Leuven  Heverlee Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiao Yin","family":"Lee","sequence":"additional","affiliation":[{"name":"Molecular Endocrinology Laboratory, Department of Cellular and Molecular Medicine KU Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tien","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Laboratory of Biomolecular Modelling and Design, Department of Chemistry KU Leuven  Heverlee Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sarah","family":"El Kharraz","sequence":"additional","affiliation":[{"name":"Molecular Endocrinology Laboratory, Department of Cellular and Molecular Medicine KU Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Florian","family":"Handle","sequence":"additional","affiliation":[{"name":"Molecular Endocrinology Laboratory, Department of Cellular and Molecular Medicine KU Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elien","family":"Smeets","sequence":"additional","affiliation":[{"name":"Molecular Endocrinology Laboratory, Department of Cellular and Molecular Medicine KU Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lisa","family":"Moris","sequence":"additional","affiliation":[{"name":"Molecular Endocrinology Laboratory, Department of Cellular and Molecular Medicine KU Leuven  Leuven Belgium"},{"name":"Department of Urology University Hospitals Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wout","family":"Devlies","sequence":"additional","affiliation":[{"name":"Molecular Endocrinology Laboratory, Department of Cellular and Molecular Medicine KU Leuven  Leuven Belgium"},{"name":"Department of Urology University Hospitals Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bram","family":"Vandewinkel","sequence":"additional","affiliation":[{"name":"Laboratory of Viral Cell Biology and Therapeutics, Department of Cellular and Molecular Medicine, Department of Microbiology, Immunology and Transplantation KU Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Irina","family":"Thiry","sequence":"additional","affiliation":[{"name":"Laboratory of Viral Cell Biology and Therapeutics, Department of Cellular and Molecular Medicine, Department of Microbiology, Immunology and Transplantation KU Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Duy Tien","family":"Ta","sequence":"additional","affiliation":[{"name":"Laboratory of Viral Cell Biology and Therapeutics, Department of Cellular and Molecular Medicine, Department of Microbiology, Immunology and Transplantation KU Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anton","family":"Gorkovskiy","sequence":"additional","affiliation":[{"name":"Laboratory of Systems Biology VIB\u2010KU Leuven Center for Microbiology  Leuven Belgium"},{"name":"Laboratory for Genetics and Genomics, Center of Microbial and Plant Genetics, Department M2S KU Leuven  Heverlee Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6397-840X","authenticated-orcid":false,"given":"Karin","family":"Voordeckers","sequence":"additional","affiliation":[{"name":"Laboratory of Systems Biology VIB\u2010KU Leuven Center for Microbiology  Leuven Belgium"},{"name":"Laboratory for Genetics and Genomics, Center of Microbial and Plant Genetics, Department M2S KU Leuven  Heverlee Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Els","family":"Henckaerts","sequence":"additional","affiliation":[{"name":"Laboratory of Viral Cell Biology and Therapeutics, Department of Cellular and Molecular Medicine, Department of Microbiology, Immunology and Transplantation KU Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vitor B.","family":"Pinheiro","sequence":"additional","affiliation":[{"name":"KU Leuven, Department of Pharmaceutical and Pharmacological Sciences Rega Institute for Medical Research  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frank","family":"Claessens","sequence":"additional","affiliation":[{"name":"Molecular Endocrinology Laboratory, Department of Cellular and Molecular Medicine KU Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin J.","family":"Verstrepen","sequence":"additional","affiliation":[{"name":"Laboratory of Systems Biology VIB\u2010KU Leuven Center for Microbiology  Leuven Belgium"},{"name":"Laboratory for Genetics and Genomics, Center of Microbial and Plant Genetics, Department M2S KU Leuven  Heverlee Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3329-2703","authenticated-orcid":false,"given":"Arnout","family":"Voet","sequence":"additional","affiliation":[{"name":"Laboratory of Biomolecular Modelling and Design, Department of Chemistry KU Leuven  Heverlee Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1048-2798","authenticated-orcid":false,"given":"Christine","family":"Helsen","sequence":"additional","affiliation":[{"name":"Molecular Endocrinology Laboratory, Department of Cellular and Molecular Medicine KU Leuven  Leuven Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,3,21]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.softx.2015.06.001"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.envint.2016.12.010"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkw408"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioeng.2004.11.002"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00894-014-2338-x"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.3389\/fendo.2022.839005"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.4103\/0976-500X.120975"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky354"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky067"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms13051"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt135"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-020-72179-0"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/yea.1744"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsbmb.2021.106043"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2011.03.008"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1089\/hgtb.2011.104"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.12792"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1997.7124"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M009870200"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aax8780"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.scitotenv.2017.11.211"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-017-02208-6"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msy225"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2006.03.014"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12934-016-0610-8"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mce.2017.05.012"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells9092077"},{"issue":"2","key":"e_1_2_8_29_1","first-page":"7","article-title":"Tamoxifen treatment for breast cancer: concept to gold standard","volume":"11","author":"Jordan VC","year":"1997","journal-title":"Oncology (Williston Park)"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1202624109"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41568-018-0001-z"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1089\/hgtb.2012.049"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-6588-5_6"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-89785-1"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11248-017-0018-1"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0050855"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gki370"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks1293"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.2179"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.1591"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrc3920"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(95)90199-x"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1016\/s1359-6446(03)02610-2"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0610155104"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0224525"},{"key":"e_1_2_8_46_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.15.6991"},{"key":"e_1_2_8_47_1","doi-asserted-by":"publisher","DOI":"10.1038\/s43586-022-00119-5"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1526-968X(200002)26:2<99::AID-GENE1>3.0.CO;2-B"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1038\/84981"},{"key":"e_1_2_8_50_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2016.09.014"},{"key":"e_1_2_8_51_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrg3927"},{"key":"e_1_2_8_52_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.100558"},{"key":"e_1_2_8_53_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.trac.2013.09.006"},{"key":"e_1_2_8_54_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms8810"},{"key":"e_1_2_8_55_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200800298"},{"key":"e_1_2_8_56_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkw1023"},{"key":"e_1_2_8_57_1","doi-asserted-by":"publisher","DOI":"10.1016\/s0092-8674(00)81717-1"},{"key":"e_1_2_8_58_1","doi-asserted-by":"publisher","DOI":"10.1186\/1754-1611-4-12"},{"key":"e_1_2_8_59_1","doi-asserted-by":"publisher","DOI":"10.1093\/jxb\/erh180"},{"key":"e_1_2_8_60_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.3696"},{"key":"e_1_2_8_61_1","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/awy126"},{"key":"e_1_2_8_62_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng.2822"},{"key":"e_1_2_8_63_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/29.12.e59"},{"key":"e_1_2_8_64_1","doi-asserted-by":"publisher","DOI":"10.1128\/mSphere.00154-18"},{"key":"e_1_2_8_65_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrev.1c00260"},{"key":"e_1_2_8_66_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.282.5387.296"},{"key":"e_1_2_8_67_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1901979116"},{"key":"e_1_2_8_68_1","doi-asserted-by":"publisher","DOI":"10.1021\/ie503060a"},{"key":"e_1_2_8_69_1","doi-asserted-by":"publisher","DOI":"10.1002\/jbm4.10450"}],"container-title":["Protein Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.4940","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,23]],"date-time":"2025-07-23T11:30:18Z","timestamp":1753270218000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pro.4940"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,21]]},"references-count":68,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2024,4]]}},"alternative-id":["10.1002\/pro.4940"],"URL":"https:\/\/doi.org\/10.1002\/pro.4940","archive":["Portico"],"relation":{},"ISSN":["0961-8368","1469-896X"],"issn-type":[{"value":"0961-8368","type":"print"},{"value":"1469-896X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,3,21]]},"assertion":[{"value":"2023-11-02","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-02-09","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-03-21","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e4940"}}