{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T08:45:30Z","timestamp":1792140330089,"version":"4.4.0"},"reference-count":50,"publisher":"American Chemical Society (ACS)","issue":"10","license":[{"start":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T00:00:00Z","timestamp":1789171200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001804","name":"Canada Research Chairs","doi-asserted-by":"publisher","award":["CERC in Glycomics"],"award-info":[{"award-number":["CERC in Glycomics"]}],"id":[{"id":"10.13039\/501100001804","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009192","name":"Alberta Innovates","doi-asserted-by":"publisher","award":["Provincial Match"],"award-info":[{"award-number":["Provincial Match"]}],"id":[{"id":"10.13039\/501100009192","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The post-transcriptional regulation of protein expression by miRNA has given rise to new technologies for therapeutic interventions and synthetic biology. To date, these technologies assume that miRNAs are repressors of protein expression; however, emerging evidence has shown that miRNA regulation of proteins is bidirectional. Although it is now clear that upregulation by miRNAs is a common event, the molecular requirements remain poorly characterized. To address this gap, we utilized the miRFluR assay, a genetically encoded dual-fluorescence reporter assay previously developed in our laboratory, to systematically explore upregulation. Using upregulation of the enzyme B3GLCT by miR-891b as a model system, we find that upregulation requires both specific sequence elements and RNA secondary structure. Mutagenesis and truncation analyses revealed that the effect is context-dependent and influenced by the local UTR structure. We also find that sites can be reengineered to be triggered by alternate miRNA, opening the door to synthetic biology manipulations. Our study provides a new perspective on upregulation by miRNA and sets the stage for further exploration of the bidirectional nature of miRNA action.<\/jats:p>","DOI":"10.1021\/acschembio.6c00503","type":"journal-article","created":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T05:01:38Z","timestamp":1789189298000},"page":"2650-2661","source":"Crossref","is-referenced-by-count":0,"title":["Elucidating the Requirements for microRNA-Mediated Upregulation Enables Reengineering of Upregulatory Sites"],"prefix":"10.1021","volume":"21","author":[{"given":"Helia Dehghan","family":"Harati","sequence":"first","affiliation":[{"name":"University of Alberta , , , ,","place":["Edmonton, Alberta, Canada, T6G 2G2"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thusini Uggalla","family":"Arachchige","sequence":"additional","affiliation":[{"name":"University of Alberta , , , ,","place":["Edmonton, Alberta, Canada, T6G 2G2"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinghai","family":"Liu","sequence":"additional","affiliation":[{"name":"University of Alberta , , , ,","place":["Edmonton, Alberta, Canada, T6G 2G2"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4791-8524","authenticated-orcid":true,"given":"Lara K.","family":"Mahal","sequence":"additional","affiliation":[{"name":"University of Alberta , , , ,","place":["Edmonton, Alberta, Canada, T6G 2G2"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,12]]},"reference":[{"issue":"8","key":"2026101523061777300_cit1","doi-asserted-by":"publisher","first-page":"629","DOI":"10.1038\/s41573-021-00219-z","article-title":"Noncoding RNA Therapeutics \u2014 Challenges and Potential Solutions","volume":"20","author":"Winkle","year":"2021","journal-title":"Nat. Rev. Drug Discov."},{"issue":"1","key":"2026101523061777300_cit2","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1016\/j.molcel.2025.11.028","article-title":"MiRNA Modules for Precise, Tunable Control of Gene Expression","volume":"86","author":"Du","year":"2026","journal-title":"Mol. Cell"},{"issue":"1","key":"2026101523061777300_cit3","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-07181-2","article-title":"Synthetic RNA-Based Logic Computation in Mammalian Cells","volume":"9","author":"Matsuura","year":"2018","journal-title":"Nat. Commun."},{"issue":"1","key":"2026101523061777300_cit4","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1089\/nat.2021.0085","article-title":"Challenges and Opportunities for Nucleic Acid Therapeutics","volume":"32","author":"Corey","year":"2022","journal-title":"Nucleic Acid Ther."},{"issue":"2","key":"2026101523061777300_cit5","doi-asserted-by":"publisher","first-page":"281","DOI":"10.1016\/S0092-8674(04)00045-5","article-title":"MicroRNAs","volume":"116","author":"Bartel","year":"2004","journal-title":"Cell"},{"key":"2026101523061777300_cit6","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.05005","article-title":"Predicting Effective MicroRNA Target Sites in Mammalian MRNAs","volume":"4","author":"Agarwal","year":"2015","journal-title":"Elife"},{"issue":"6472","key":"2026101523061777300_cit7","doi-asserted-by":"publisher","DOI":"10.1126\/science.aav1741","article-title":"The Biochemical Basis of MicroRNA Targeting Efficacy","volume":"366","author":"McGeary","year":"2019","journal-title":"Science (1979)"},{"issue":"10","key":"2026101523061777300_cit8","doi-asserted-by":"publisher","first-page":"1900","DOI":"10.1021\/acschembio.1c00247","article-title":"High-Throughput MiRFluR Platform Identifies MiRNA Regulating B3GLCT That Predict Peters\u2019 Plus Syndrome Phenotype, Supporting the MiRNA Proxy Hypothesis","volume":"16","author":"Thu","year":"2021","journal-title":"ACS Chem. Biol."},{"key":"2026101523061777300_cit9","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2026.113217","article-title":"MicroRNAs Bidirectionally Regulate FUT1 to Modulate \u03b1-1,2-Fucosylation and Cancer-Associated Biology","volume-title":"J. Biol. Chem.","author":"Batu","year":"2026"},{"issue":"11","key":"2026101523061777300_cit10","doi-asserted-by":"publisher","first-page":"1527","DOI":"10.1021\/acscentsci.2c00748","article-title":"High-Throughput Analysis Reveals MiRNA Upregulating \u03b1-2,6-Sialic Acid through Direct MiRNA\u2212MRNA Interactions","volume":"8","author":"Jame-Chenarboo","year":"2022","journal-title":"ACS Cent. Sci."},{"issue":"2","key":"2026101523061777300_cit11","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adr0277","article-title":"Screening the Human MiRNA Interactome Reveals Coordinated Up-Regulation in Melanoma, Adding Bidirectional Regulation to MiRNA Networks","volume":"11","author":"Jame-Chenarboo","year":"2025","journal-title":"Sci. Adv."},{"issue":"4","key":"2026101523061777300_cit12","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2025.108340","article-title":"Profiling the Regulatory Landscape of Sialylation through MiRNA Targeting of CMP- Sialic Acid Synthetase","volume":"301","author":"Jame-Chenarboo","year":"2025","journal-title":"J. Biol. Chem."},{"issue":"9","key":"2026101523061777300_cit13","doi-asserted-by":"publisher","first-page":"2159","DOI":"10.1021\/acschembio.5c00296","article-title":"MicroRNAs Regulate Cellular Magnesium by Tuning Expression of the Plasma Membrane Protein CNNM4","volume":"20","author":"Lazarou","year":"2025","journal-title":"ACS Chem. Biol."},{"issue":"5858","key":"2026101523061777300_cit14","doi-asserted-by":"publisher","first-page":"1931","DOI":"10.1126\/science.1149460","article-title":"Switching from Repression to Activation: MicroRNAs Can Up-Regulate Translation","volume":"318","author":"Vasudevan","year":"2007","journal-title":"Science"},{"issue":"7256","key":"2026101523061777300_cit15","doi-asserted-by":"publisher","first-page":"705","DOI":"10.1038\/nature08195","article-title":"MiR-145 and MiR-143 Regulate Smooth Muscle Cell Fate and Plasticity","volume":"460","author":"Cordes","year":"2009","journal-title":"Nature"},{"issue":"3","key":"2026101523061777300_cit16","doi-asserted-by":"publisher","first-page":"607","DOI":"10.1016\/j.cell.2014.05.047","article-title":"MicroRNA Directly Enhances Mitochondrial Translation during Muscle Differentiation","volume":"158","author":"Zhang","year":"2014","journal-title":"Cell"},{"issue":"11","key":"2026101523061777300_cit17","doi-asserted-by":"publisher","first-page":"1640","DOI":"10.1261\/rna.2191905","article-title":"A Crucial Role for GW182 and the DCP1:DCP2 Decapping Complex in MiRNA-Mediated Gene Silencing","volume":"11","author":"Rehwinkel","year":"2005","journal-title":"RNA"},{"issue":"27","key":"2026101523061777300_cit18","doi-asserted-by":"publisher","first-page":"17897","DOI":"10.1074\/jbc.R900012200","article-title":"The RNA-Induced Silencing Complex: A Versatile Gene-Silencing Machine","volume":"284","author":"Pratt","year":"2009","journal-title":"J. Biol. Chem."},{"issue":"4","key":"2026101523061777300_cit19","doi-asserted-by":"publisher","first-page":"3973","DOI":"10.18632\/aging.205571","article-title":"MicroRNA-320a Enhances LRWD1 Expression through the AGO2\/FXR1-Dependent Pathway to Affect Cell Behaviors and the Oxidative Stress Response in Human Testicular Embryonic Carcinoma Cells","volume":"16","author":"Lin","year":"2024","journal-title":"Aging"},{"issue":"13","key":"2026101523061777300_cit20","doi-asserted-by":"publisher","first-page":"7439","DOI":"10.1093\/nar\/gkaa478","article-title":"Argonaute Binding within 3\u2032-Untranslated Regions Poorly Predicts Gene Repression","volume":"48","author":"Chu","year":"2020","journal-title":"Nucleic Acids Res."},{"issue":"5","key":"2026101523061777300_cit21","doi-asserted-by":"publisher","first-page":"375","DOI":"10.14348\/molcells.2016.0013","article-title":"MicroRNA Target Recognition: Insights from Transcriptome-Wide Non-Canonical Interactions","volume":"39","author":"Seok","year":"2016","journal-title":"Mol. Cells"},{"issue":"4","key":"2026101523061777300_cit22","doi-asserted-by":"publisher","first-page":"741","DOI":"10.1016\/j.molcel.2019.06.012","article-title":"High-Throughput Analysis Reveals Rules for Target RNA Binding and Cleavage by AGO2","volume":"75","author":"Becker","year":"2019","journal-title":"Mol. Cell"},{"issue":"1","key":"2026101523061777300_cit23","doi-asserted-by":"publisher","DOI":"10.1186\/s12859-021-04164-x","article-title":"Comprehensive Machine-Learning-Based Analysis of MicroRNA\u2212Target Interactions Reveals Variable Transferability of Interaction Rules across Species","volume":"22","author":"Ben Or","year":"2021","journal-title":"BMC Bioinformatics"},{"issue":"3","key":"2026101523061777300_cit24","doi-asserted-by":"publisher","first-page":"654","DOI":"10.1016\/j.cell.2013.03.043","article-title":"Mapping the Human MiRNA Interactome by CLASH Reveals Frequent Noncanonical Binding","volume":"153","author":"Helwak","year":"2013","journal-title":"Cell"},{"issue":"4","key":"2026101523061777300_cit25","doi-asserted-by":"publisher","first-page":"669","DOI":"10.1158\/1541-7786.MCR-17-0321","article-title":"Characterization and Evidence of the MiR-888 Cluster as a Novel Cancer Network in Prostate","volume":"16","author":"Hasegawa","year":"2018","journal-title":"Mol. Cancer Res."},{"issue":"1","key":"2026101523061777300_cit26","doi-asserted-by":"publisher","DOI":"10.1186\/s13046-021-01910-0","article-title":"ChrXq273 MiRNA Cluster Functions in Cancer Development","volume":"40","author":"Yoshida","year":"2021","journal-title":"J. Exp. Clin. Cancer Res."},{"issue":"Web Server","key":"2026101523061777300_cit27","doi-asserted-by":"publisher","first-page":"W451","DOI":"10.1093\/nar\/gkl243","article-title":"RNAhybrid: MicroRNA Target Prediction Easy, Fast and Flexible","volume":"34","author":"Kruger","year":"2006","journal-title":"Nucleic Acids Res."},{"issue":"10","key":"2026101523061777300_cit28","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0206239","article-title":"MiRWalk: An Online Resource for Prediction of MicroRNA Binding Sites","volume":"13","author":"Sticht","year":"2018","journal-title":"PLoS One"},{"issue":"7","key":"2026101523061777300_cit29","doi-asserted-by":"publisher","first-page":"1297","DOI":"10.1261\/rna.1082708","article-title":"Molecular Architecture of a MiRNA-Regulated 3\u2032 UTR","volume":"14","author":"Didiano","year":"2008","journal-title":"RNA"},{"issue":"2","key":"2026101523061777300_cit30","doi-asserted-by":"publisher","first-page":"471","DOI":"10.1016\/j.celrep.2013.08.050","article-title":"An Extensive Network of TET2-Targeting MicroRNAs Regulates Malignant Hematopoiesis","volume":"5","author":"Cheng","year":"2013","journal-title":"Cell Rep."},{"issue":"7","key":"2026101523061777300_cit31","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0068204","article-title":"Large-Scale Screens of MiRNA-MRNA Interactions Unveiled That the 3\u2032UTR of a Gene Is Targeted by Multiple MiRNAs","volume":"8","author":"Zhou","year":"2013","journal-title":"PLoS One"},{"issue":"10","key":"2026101523061777300_cit32","doi-asserted-by":"publisher","first-page":"1751","DOI":"10.1136\/gutjnl-2017-315318","article-title":"MicroRNA-92a-1\u22125p Increases CDX2 by Targeting FOXD1 in Bile Acids-Induced Gastric Intestinal Metaplasia","volume":"68","author":"Li","year":"2019","journal-title":"Gut"},{"issue":"50","key":"2026101523061777300_cit33","doi-asserted-by":"publisher","first-page":"82338","DOI":"10.18632\/oncotarget.11001","article-title":"MicroRNA-891b Is an Independent Prognostic Factor of Pancreatic Cancer by Targeting Cbl-b to Suppress the Growth of Pancreatic Cancer Cells","volume":"7","author":"Dong","year":"2016","journal-title":"Oncotarget"},{"issue":"1","key":"2026101523061777300_cit34","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-43065-w","article-title":"Rational Design of MicroRNA-Responsive Switch for Programmable Translational Control in Mammalian Cells","volume":"14","author":"Ning","year":"2023","journal-title":"Nat. Commun."},{"issue":"3","key":"2026101523061777300_cit35","doi-asserted-by":"publisher","first-page":"671","DOI":"10.1093\/molbev\/msp284","article-title":"Evolution of an X-Linked Primate-Specific Micro RNA Cluster","volume":"27","author":"Li","year":"2010","journal-title":"Mol. Biol. Evol."},{"issue":"3","key":"2026101523061777300_cit36","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1002\/wrna.121","article-title":"Posttranscriptional Upregulation by MicroRNAs","volume":"3","author":"Vasudevan","year":"2012","journal-title":"WIREs RNA"},{"key":"2026101523061777300_cit37","doi-asserted-by":"publisher","DOI":"10.3389\/fgene.2013.00008","article-title":"Role of MiRNAs in CD4 T Cell Plasticity during Inflammation and Tolerance","volume":"4","author":"Sethi","year":"2013","journal-title":"Front. Genet."},{"key":"2026101523061777300_cit38","doi-asserted-by":"publisher","DOI":"10.1155\/2014\/970607","article-title":"Mechanisms of MiRNA-Mediated Gene Regulation from Common Downregulation to MRNA-Specific Upregulation","volume":"2014","author":"Valinezhad Orang","year":"2014","journal-title":"Int. J. Genomics"},{"issue":"1","key":"2026101523061777300_cit39","doi-asserted-by":"publisher","DOI":"10.1186\/1758-907X-1-11","article-title":"How Do MiRNAs Mediate Translational Repression?","volume":"1","author":"Gu","year":"2010","journal-title":"Silence"},{"issue":"1","key":"2026101523061777300_cit40","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1261\/rna.034769.112","article-title":"Human TNRC6A Is an Argonaute-Navigator Protein for MicroRNA-Mediated Gene Silencing in the Nucleus","volume":"19","author":"Nishi","year":"2013","journal-title":"RNA"},{"key":"2026101523061777300_cit41","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.12880","article-title":"Cotranslational MicroRNA Mediated Messenger RNA Destabilization","volume":"5","author":"Tat","year":"2016","journal-title":"Elife"},{"issue":"4","key":"2026101523061777300_cit42","doi-asserted-by":"publisher","first-page":"533","DOI":"10.1016\/j.molcel.2006.01.031","article-title":"Short RNAs Repress Translation after Initiation in Mammalian Cells","volume":"21","author":"Petersen","year":"2006","journal-title":"Mol. Cell"},{"issue":"1","key":"2026101523061777300_cit43","doi-asserted-by":"publisher","first-page":"586","DOI":"10.1093\/nar\/gks1076","article-title":"MiRNA Repression of Translation in Vitro Takes Place during 43S Ribosomal Scanning","volume":"41","author":"Ricci","year":"2013","journal-title":"Nucleic Acids Res."},{"issue":"18","key":"2026101523061777300_cit44","doi-asserted-by":"publisher","first-page":"8692","DOI":"10.1093\/nar\/gkt619","article-title":"MiRISC Recruits Decapping Factors to MiRNA Targets to Enhance Their Degradation","volume":"41","author":"Nishihara","year":"2013","journal-title":"Nucleic Acids Res."},{"issue":"11","key":"2026101523061777300_cit45","doi-asserted-by":"publisher","first-page":"1211","DOI":"10.1038\/nsmb.2149","article-title":"MiRNA-Mediated Deadenylation Is Orchestrated by GW182 through Two Conserved Motifs That Interact with CCR4\u2212NOT","volume":"18","author":"Fabian","year":"2011","journal-title":"Nat. Struct. Mol. Biol."},{"issue":"6209","key":"2026101523061777300_cit46","doi-asserted-by":"publisher","first-page":"608","DOI":"10.1126\/science.1258040","article-title":"Structural Basis for MicroRNA Targeting","volume":"346","author":"Schirle","year":"2014","journal-title":"Science"},{"issue":"1","key":"2026101523061777300_cit47","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-27177-9","article-title":"A Structured RNA Motif Locks Argonaute2:MiR-122 onto the 5\u2032 End of the HCV Genome","volume":"12","author":"Gebert","year":"2021","journal-title":"Nat. Commun."},{"issue":"5","key":"2026101523061777300_cit48","doi-asserted-by":"publisher","first-page":"285","DOI":"10.1089\/nat.2018.0734","article-title":"MicroRNAs Enable MRNA Therapeutics to Selectively Program Cancer Cells to Self-Destruct","volume":"28","author":"Jain","year":"2018","journal-title":"Nucleic Acid Ther."},{"issue":"12","key":"2026101523061777300_cit49","doi-asserted-by":"publisher","DOI":"10.3390\/cells8121540","article-title":"From Endogenous to Synthetic MicroRNA-Mediated Regulatory Circuits: An Overview","volume":"8","author":"Ferro","year":"2019","journal-title":"Cells"},{"issue":"2","key":"2026101523061777300_cit50","doi-asserted-by":"publisher","first-page":"234","DOI":"10.1101\/gr.205146.116","article-title":"MicroRNA Target Prediction Programs Predict Many False Positives","volume":"27","author":"Pinz\u00f3n","year":"2017","journal-title":"Genome Res."}],"container-title":["ACS Chemical Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acschembio.6c00503","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/21\/10\/2650\/68490476\/cb-2026-00503e.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/21\/10\/2650\/68490476\/cb-2026-00503e.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T08:10:26Z","timestamp":1792138226000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acbcct\/article\/21\/10\/2650\/5428806\/Elucidating-the-Requirements-for-microRNA-Mediated"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,12]]},"references-count":50,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2026,9,12]]},"published-print":{"date-parts":[[2026,10,16]]}},"URL":"https:\/\/doi.org\/10.1021\/acschembio.6c00503","relation":{},"ISSN":["1554-8929","1554-8937"],"issn-type":[{"value":"1554-8929","type":"print"},{"value":"1554-8937","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,12]]}}}