{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T14:08:04Z","timestamp":1789567684113,"version":"build-2803163510"},"reference-count":46,"publisher":"American Chemical Society (ACS)","issue":"36","license":[{"start":{"date-parts":[[2026,9,6]],"date-time":"2026-09-06T00:00:00Z","timestamp":1788652800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,9,6]],"date-time":"2026-09-06T00:00:00Z","timestamp":1788652800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,9,6]],"date-time":"2026-09-06T00:00:00Z","timestamp":1788652800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2025YFA0921100, 2024YFA0917100"],"award-info":[{"award-number":["2025YFA0921100, 2024YFA0917100"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Shanghai Municipal Science and Technology Major Project"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Yarrowia lipolytica is a promising industrial host, yet its metabolic engineering potential remains limited by insufficient genetic tools. Here, we engineered a synthetic Transcription Activation Toolkit (TAT) based on LacI\u2013VPRH, a chimeric protein fusing the prokaryotic DNA-binding domain LacI with the eukaryotic activation domain VPRH. Systematic optimization of LacO copy numbers and core promoter composition achieved up to 205-fold gene activation. The TAT platform was further expanded to construct bidirectional expression systems and enable multiplexed gene control. Applied to resveratrol biosynthesis via a \u201cpush-pull\u201d strategy, CRISPR\/Cas9-mediated integration of TAT-controlled synthetic promoters upregulated the shikimate pathway genes aroM10 and aroC alongside the rate-limiting enzyme ST1, achieving a shake-flask titer of 2.715 g\/L\u2500the highest reported to date. Additionally, an IPTG-inducible \u201cturn-on\u201d system (TAT-2.0) incorporating the antiLacI9 mutant was developed for small-molecule-responsive transcriptional control. Collectively, the modular TAT system provides a versatile strategy for precise metabolic pathway optimization in Y. lipolytica.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c05415","type":"journal-article","created":{"date-parts":[[2026,9,6]],"date-time":"2026-09-06T15:01:37Z","timestamp":1788706897000},"page":"28570-28582","source":"Crossref","is-referenced-by-count":0,"title":["Engineering a Transcription\nActivation Toolkit by\nRewiring Prokaryotic Transcription Factors and Synthetic Yeast Promoters\nin\n                    <i>Yarrowia lipolytica<\/i>"],"prefix":"10.1021","volume":"74","author":[{"given":"Yunhe","family":"Li","sequence":"first","affiliation":[{"name":"East China University of Science and Technology , , ,","place":["Shanghai, China, 200237"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-0748-4828","authenticated-orcid":true,"given":"Wenping","family":"Wei","sequence":"additional","affiliation":[{"name":"Zhejiang University of Technology , , , ,","place":["Hangzhou, Zhejiang, China, 310014"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ping","family":"Zhang","sequence":"additional","affiliation":[{"name":"East China University of Science and Technology , , ,","place":["Shanghai, China, 200237"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting","family":"Huang","sequence":"additional","affiliation":[{"name":"Zhejiang University of Technology , , , ,","place":["Hangzhou, Zhejiang, China, 310014"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanzhe","family":"Shang","sequence":"additional","affiliation":[{"name":"East China University of Science and Technology , , ,","place":["Shanghai, China, 200237"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9241-1293","authenticated-orcid":true,"given":"Ying","family":"Zhou","sequence":"additional","affiliation":[{"name":"East China University of Science and Technology , , ,","place":["Shanghai, China, 200237"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5555-5359","authenticated-orcid":true,"given":"Bang-Ce","family":"Ye","sequence":"additional","affiliation":[{"name":"East China University of Science and Technology , , ,","place":["Shanghai, China, 200237"]},{"name":"Zhejiang University of Technology , , , ,","place":["Hangzhou, Zhejiang, China, 310014"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,6]]},"reference":[{"issue":"8","key":"2026091609084736800_cit1","doi-asserted-by":"publisher","first-page":"1781","DOI":"10.1021\/jacsau.2c00344","article-title":"Designing\nMicrobial Cell Factories for the Production of Chemicals","volume":"2","author":"Cho","year":"2022","journal-title":"JACS Au"},{"issue":"10","key":"2026091609084736800_cit2","doi-asserted-by":"publisher","first-page":"1085","DOI":"10.1016\/j.tibtech.2018.05.004","article-title":"Synthetic Biology\nExpands the Industrial Potential\nof Yarrowia lipolytica","volume":"36","author":"Markham","year":"2018","journal-title":"Trends\nBiotechnol."},{"issue":"2","key":"2026091609084736800_cit3","doi-asserted-by":"publisher","first-page":"242","DOI":"10.1016\/j.tibtech.2022.07.006","article-title":"What Makes Yarrowia lipolytica Well\nSuited for Industry?","volume":"41","author":"Park","year":"2023","journal-title":"Trends Biotechnol."},{"key":"2026091609084736800_cit4","doi-asserted-by":"publisher","first-page":"65","DOI":"10.3389\/fbioe.2017.00065","article-title":"Integrating\nCellular and Bioprocess Engineering in the Non-Conventional Yeast\nYarrowia lipolytica for Biodiesel Production: A Review","volume":"5","author":"Xie","year":"2017","journal-title":"Front. Bioeng. Biotechnol."},{"issue":"6","key":"2026091609084736800_cit5","doi-asserted-by":"publisher","first-page":"423","DOI":"10.1002\/elsc.201800147","article-title":"Cellular and Metabolic\nEngineering of Oleaginous Yeast Yarrowia lipolytica for Bioconversion\nof Hydrophobic Substrates into High-value Products","volume":"19","author":"Soong","year":"2019","journal-title":"Eng. Life Sci."},{"issue":"3","key":"2026091609084736800_cit6","doi-asserted-by":"publisher","first-page":"639","DOI":"10.1021\/acssynbio.2c00569","article-title":"Metabolic\nEngineering of Yarrowia lipolytica for Terpenoid\nProduction: Tools and\nStrategies","volume":"12","author":"Zhang","year":"2023","journal-title":"ACS Synth. Biol."},{"key":"2026091609084736800_cit7","doi-asserted-by":"publisher","first-page":"129129","DOI":"10.1016\/j.biortech.2023.129129","article-title":"Metabolic\nEngineering for the High-Yield Production of Polydatin in Yarrowia lipolytica","volume":"381","author":"Shang","year":"2023","journal-title":"Bioresour. Technol."},{"key":"2026091609084736800_cit8","doi-asserted-by":"publisher","first-page":"123726","DOI":"10.1016\/j.biortech.2020.123726","article-title":"Metabolically\nEngineering of Yarrowia lipolytica for the Biosynthesis\nof Naringenin from a Mixture of Glucose and Xylose","volume":"314","author":"Wei","year":"2020","journal-title":"Bioresour. Technol."},{"key":"2026091609084736800_cit9","doi-asserted-by":"publisher","first-page":"129421","DOI":"10.1016\/j.biortech.2023.129421","article-title":"Metabolic Engineering of Yarrowia lipolytica for High-Level Production of Scutellarin","volume":"385","author":"Zhang","year":"2023","journal-title":"Bioresour. Technol."},{"issue":"5","key":"2026091609084736800_cit10","doi-asserted-by":"publisher","first-page":"1364","DOI":"10.1021\/acs.jafc.9b07151","article-title":"Engineering Yarrowia lipolytica for Enhanced Production of Arbutin","volume":"68","author":"Shang","year":"2020","journal-title":"J. Agric.\nFood Chem."},{"issue":"8","key":"2026091609084736800_cit11","doi-asserted-by":"publisher","first-page":"2223","DOI":"10.1111\/1751-7915.14060","article-title":"Identification\nof Genome Integration Sites for Developing a CRISPR-based Gene Expression\nToolkit in Yarrowia lipolytica","volume":"15","author":"Liu","year":"2022","journal-title":"Microb. Biotechnol."},{"issue":"1","key":"2026091609084736800_cit12","doi-asserted-by":"publisher","first-page":"149","DOI":"10.1186\/s13068-021-02002-z","article-title":"Hybrid Promoter Engineering\nStrategies in Yarrowia lipolytica: Isoamyl Alcohol\nProduction as a Test Study","volume":"14","author":"Zhao","year":"2021","journal-title":"Biotechnol. Biofuels"},{"issue":"8","key":"2026091609084736800_cit13","doi-asserted-by":"publisher","DOI":"10.1111\/1751-7915.70185","article-title":"Unlocking the Synthetic\nPotential of Yarrowia lipolytica: Innovating Gene\nExpression Tools","volume":"18","author":"Liu","year":"2025","journal-title":"Microb.\nBiotechnol."},{"issue":"14","key":"2026091609084736800_cit14","doi-asserted-by":"publisher","first-page":"1267","DOI":"10.1002\/(SICI)1097-0061(1998100)14:14&lt;1267::AID-YEA327&gt;3.0.CO;2-2","article-title":"Comparison\nof Expression Systems in the Yeasts Saccharomyces Cerevisiae, Hansenula\nPolymorpha, Klyveromyces Lactis, Schizosaccharomyces Pombe and Yarrowia\nlipolytica. Cloning of two novel promoters from Yarrowia lipolytica","volume":"14","author":"Mu\u0308ller","year":"1998","journal-title":"Yeast"},{"key":"2026091609084736800_cit15","doi-asserted-by":"publisher","first-page":"68","DOI":"10.1016\/j.meteno.2017.09.001","article-title":"YaliBricks, a Versatile Genetic Toolkit\nfor Streamlined and Rapid\nPathway Engineering in Yarrowia lipolytica","volume":"5","author":"Wong","year":"2017","journal-title":"Metab. Eng. Commun."},{"issue":"2","key":"2026091609084736800_cit16","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1002\/yea.1917","article-title":"Engineering Yarrowia lipolytica to\nExpress Secretory Invertase with Strong FBA1IN Promoter","volume":"29","author":"Hong","year":"2012","journal-title":"Yeast"},{"issue":"10","key":"2026091609084736800_cit17","doi-asserted-by":"publisher","first-page":"4621","DOI":"10.1128\/jb.169.10.4621-4629.1987","article-title":"Cloning\nand Sequencing of the Alkaline Extracellular\nProtease Gene of Yarrowia lipolytica","volume":"169","author":"Davidow","year":"1987","journal-title":"J. Bacteriol."},{"key":"2026091609084736800_cit18","doi-asserted-by":"publisher","first-page":"132656","DOI":"10.1016\/j.biortech.2025.132656","article-title":"Expanding\nthe Genetic Toolkit of Yarrowia lipolytica: Dynamic\nPromoter Engineering Enables High-Titer Biosynthesis of 3-Hydroxypropionic\nAcid","volume":"432","author":"Kang","year":"2025","journal-title":"Bioresour. Technol."},{"key":"2026091609084736800_cit19","doi-asserted-by":"publisher","first-page":"7905","DOI":"10.1128\/AEM.05763-11","article-title":"Tuning Gene Expression in Yarrowia\nlipolytica by a Hybrid Promoter Approach","volume":"77","author":"Blazeck","year":"2011","journal-title":"Appl.\nEnviron. Microbiol."},{"issue":"1","key":"2026091609084736800_cit20","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1016\/j.jbiotec.2003.10.027","article-title":"Heterologous Protein Expression and\nSecretion in the Non-Conventional Yeast Yarrowia lipolytica: A Review","volume":"109","author":"Madzak","year":"2004","journal-title":"J. Biotechnol."},{"issue":"1","key":"2026091609084736800_cit21","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1186\/s12934-017-0647-3","article-title":"Using a Vector Pool\nContaining Variable-Strength Promoters\nto Optimize Protein Production in Yarrowia lipolytica","volume":"16","author":"Dulermo","year":"2017","journal-title":"Microb. Cell Fact."},{"key":"2026091609084736800_cit22","doi-asserted-by":"publisher","first-page":"foaf023","DOI":"10.1093\/femsyr\/foaf023","article-title":"Global Transcription\nMachinery Engineering in Yarrowia lipolytica","volume":"25","author":"Celin\u0301ska","year":"2025","journal-title":"FEMS Yeast Res."},{"issue":"6","key":"2026091609084736800_cit23","doi-asserted-by":"publisher","DOI":"10.1128\/msphere.00450-22","article-title":"The Zinc Finger\nTranscription Factor Fts2 Represses the Yeast-to-Filament Transition\nin the Dimorphic Yeast Yarrowia lipolytica","volume":"7","author":"Chen","year":"2022","journal-title":"mSphere"},{"issue":"5","key":"2026091609084736800_cit24","doi-asserted-by":"publisher","first-page":"foy037","DOI":"10.1093\/femsyr\/foy037","article-title":"Overexpression\nScreen Reveals Transcription\nFactors Involved in Lipid Accumulation in Yarrowia lipolytica","volume":"18","author":"Leplat","year":"2018","journal-title":"FEMS Yeast Res."},{"issue":"23","key":"2026091609084736800_cit25","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2426686122","article-title":"High-Throughput\nMetabolic Engineering\nof Yarrowia lipolytica through Gene Expression Tuning","volume":"122","author":"Jiang","year":"2025","journal-title":"Proc. Natl. Acad. Sci. U.S. A."},{"issue":"1","key":"2026091609084736800_cit26","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1186\/s13068-018-1114-z","article-title":"Engineering Global Transcription\nto Tune Lipophilic Properties in Yarrowia lipolytica","volume":"11","author":"Wang","year":"2018","journal-title":"Biotechnol. Biofuels"},{"key":"2026091609084736800_cit27","doi-asserted-by":"publisher","first-page":"1198","DOI":"10.1016\/j.tibtech.2025.10.014","article-title":"Dynamic\nRegulation of Oleaginous\nYeast Metabolism for Chemical Production","volume":"44","author":"Wang","year":"2026","journal-title":"Trends\nBiotechnol."},{"issue":"47","key":"2026091609084736800_cit28","doi-asserted-by":"publisher","first-page":"30380","DOI":"10.1021\/acs.jafc.5c07541","article-title":"Systematic Metabolic Engineering\nof Oleaginous Yeast Yarrowia lipolytica for Efficient Synthesis of Polyketide 6-Methylsalicylic\nAcid","volume":"73","author":"Wei","year":"2025","journal-title":"J. Agric. Food Chem."},{"issue":"5","key":"2026091609084736800_cit29","doi-asserted-by":"publisher","first-page":"1022","DOI":"10.1021\/acssynbio.0c00122","article-title":"Engineering Prokaryotic\nTranscriptional Activator XylR\nas a Xylose-Inducible Biosensor for Transcription Activation in Yeast","volume":"9","author":"Wei","year":"2020","journal-title":"ACS Synth. Biol."},{"key":"2026091609084736800_cit30","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1016\/j.ymben.2020.05.005","article-title":"Coupling Metabolic\nAddiction with Negative Autoregulation to Improve Strain Stability\nand Pathway Yield","volume":"61","author":"Lv","year":"2020","journal-title":"Metab. Eng."},{"key":"2026091609084736800_cit31","doi-asserted-by":"publisher","first-page":"318","DOI":"10.1016\/S0022-2836(61)80072-7","article-title":"Genetic Regulatory\nMechanisms in the Synthesis of Proteins","volume":"3","author":"Jacob","year":"1961","journal-title":"J.\nMol. Biol."},{"issue":"6","key":"2026091609084736800_cit32","doi-asserted-by":"publisher","first-page":"1891","DOI":"10.1073\/pnas.56.6.1891","article-title":"Isolation of the Lac Repressor","volume":"56","author":"Gilbert","year":"1966","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"issue":"3","key":"2026091609084736800_cit33","doi-asserted-by":"publisher","first-page":"401","DOI":"10.1016\/0022-2836(70)90074-4","article-title":"The lac represser-operator\ninteraction","volume":"53","author":"Riggs","year":"1970","journal-title":"J. Mol. Biol."},{"issue":"5673","key":"2026091609084736800_cit34","doi-asserted-by":"publisher","first-page":"765","DOI":"10.1038\/274765a0","article-title":"Sequence\nof the lacI Gene","volume":"274","author":"Farabaugh","year":"1978","journal-title":"Nature"},{"issue":"5253","key":"2026091609084736800_cit35","doi-asserted-by":"publisher","first-page":"1247","DOI":"10.1126\/science.271.5253.1247","article-title":"Crystal Structure of\nthe Lactose Operon Repressor and Its Complexes with DNA and Inducer","volume":"271","author":"Lewis","year":"1996","journal-title":"Science"},{"issue":"17","key":"2026091609084736800_cit36","doi-asserted-by":"publisher","first-page":"10187","DOI":"10.1093\/nar\/gkac765","article-title":"Yeast Transcriptional\nDevice Libraries Enable Precise Synthesis of Value-Added Chemicals\nfrom Methanol","volume":"50","author":"Zhu","year":"2022","journal-title":"Nucleic Acids Res."},{"issue":"12","key":"2026091609084736800_cit37","doi-asserted-by":"publisher","first-page":"3912","DOI":"10.1021\/acssynbio.2c00423","article-title":"A Toolkit for Precise, Multigene\nControl in Saccharomyces Cerevisiae","volume":"11","author":"Sanford","year":"2022","journal-title":"ACS Synth. Biol."},{"issue":"9","key":"2026091609084736800_cit38","doi-asserted-by":"publisher","DOI":"10.1002\/biot.201700584","article-title":"Multiplexed CRISPR\nActivation of Cryptic Sugar Metabolism Enables Yarrowia lipolytica Growth on Cellobiose","volume":"13","author":"Schwartz","year":"2018","journal-title":"Biotechnol. J."},{"issue":"6","key":"2026091609084736800_cit39","doi-asserted-by":"publisher","first-page":"294","DOI":"10.1016\/S0968-0004(03)00088-4","article-title":"The Herpes Simplex Virus VP16-Induced Complex: The Makings of a Regulatory\nSwitch","volume":"28","author":"Wysocka","year":"2003","journal-title":"Trends Biochem. Sci."},{"key":"2026091609084736800_cit40","doi-asserted-by":"publisher","first-page":"5","DOI":"10.3389\/fbioe.2017.00063","article-title":"Synthetic\nPromoters and Transcription Factors for Heterologous Protein Expression\nin Saccharomyces Cerevisiae","volume":"5","author":"Machens","year":"2017","journal-title":"Front. Bioeng.\nBiotechnol."},{"issue":"6","key":"2026091609084736800_cit41","doi-asserted-by":"publisher","first-page":"1790","DOI":"10.1111\/j.1365-2958.2006.05055.x","article-title":"The UGA3-GLT1\nIntergenic Region Constitutes a Promoter\nWhose Bidirectional Nature Is Determined by Chromatin Organization\nin Saccharomyces Cerevisiae","volume":"59","author":"Ishida","year":"2006","journal-title":"Mol. Microbiol."},{"issue":"1","key":"2026091609084736800_cit42","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1186\/s12934-023-02020-6","article-title":"Bidirectional\nHybrid Erythritol-Inducible Promoter for Synthetic Biology in Yarrowia lipolytica","volume":"22","author":"Vidal","year":"2023","journal-title":"Microb. Cell\nFact."},{"issue":"1","key":"2026091609084736800_cit43","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks972","article-title":"Rationally Designed Bidirectional\nPromoter Improves the Evolutionary Stability of Synthetic Genetic\nCircuits","volume":"41","author":"Yang","year":"2013","journal-title":"Nucleic Acids Res."},{"issue":"1","key":"2026091609084736800_cit44","doi-asserted-by":"publisher","first-page":"4976","DOI":"10.1038\/s41467-019-12961-5","article-title":"Rewiring Carbon Metabolism in Yeast for High Level Production of\nAromatic Chemicals","volume":"10","author":"Liu","year":"2019","journal-title":"Nat. Commun."},{"issue":"7","key":"2026091609084736800_cit45","doi-asserted-by":"publisher","first-page":"3167","DOI":"10.1007\/s00253-019-09664-8","article-title":"Understanding Lipogenesis\nby Dynamically Profiling Transcriptional Activity of Lipogenic Promoters\nin Yarrowia lipolytica","volume":"103","author":"Liu","year":"2019","journal-title":"Appl.\nMicrobiol. Biotechnol."},{"issue":"1","key":"2026091609084736800_cit46","doi-asserted-by":"publisher","first-page":"6","DOI":"10.1021\/acssynbio.6b00048","article-title":"Fourteen\nWays to Reroute Cooperative\nCommunication in the Lactose Repressor: Engineering Regulatory Proteins\nwith Alternate Repressive Functions","volume":"6","author":"Richards","year":"2017","journal-title":"ACS Synth.\nBiol."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c05415","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/36\/28570\/68396331\/acs.jafc.6c05415.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/36\/28570\/68396331\/acs.jafc.6c05415.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T13:33:37Z","timestamp":1789565617000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/36\/28570\/5421917\/Engineering-a-Transcription-Activation-Toolkit-by"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,6]]},"references-count":46,"journal-issue":{"issue":"36","published-online":{"date-parts":[[2026,9,6]]},"published-print":{"date-parts":[[2026,9,16]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c05415","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,6]]}}}