{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,18]],"date-time":"2026-10-18T18:27:13Z","timestamp":1792348033143,"version":"4.4.0"},"reference-count":58,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2025,9,17]],"date-time":"2025-09-17T00:00:00Z","timestamp":1758067200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Shanghai Jiao Tong University Transmed Awards Program","award":["20190104"],"award-info":[{"award-number":["20190104"]}]},{"DOI":"10.13039\/501100012166","name":"National Key R&D Program of China","doi-asserted-by":"publisher","award":["2018YFA0900600"],"award-info":[{"award-number":["2018YFA0900600"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Sustainable Utilization Capacity Building Project for Valuable Chinese Medicinal Resources","award":["2060302"],"award-info":[{"award-number":["2060302"]}]},{"name":"Scientific and Technological Project of China Academy of Chinese Medical Sciences","award":["CI2024C005YN, CI2023E002"],"award-info":[{"award-number":["CI2024C005YN, CI2023E002"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,1,29]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Tanshinone accumulation serves as a critical determinant of medicinal value in Salvia miltiorrhiza cultivars. Precise fine-tuning of tanshinone biosynthesis while preserving elite agronomic traits remains a pivotal challenge in molecular breeding. Here, we report, for the first time, the successful application of CRISPR\/Cas9-mediated upstream open reading frame (uORF) editing in medicinal plants to enhance the production of specialized metabolites. Five evolutionarily conserved uORFs identified in the 5\u2032 leader sequence of the key diterpene synthase gene SmCPS1 were strategically edited to modulate post-transcriptional regulation. Homozygous mutants engineered through precision gene editing exhibited 1.19- to 1.81-fold enhanced tanshinone accumulation compared to the controls, correlating with coordinated transcriptional activation of core biosynthetic genes (SmHMGR1, SmKSL1, SmCYP76AH1, SmCYP76AH3). Integrative molecular analyses demonstrated unchanged SmCPS1 transcript levels and enhanced protein accumulation, mechanistically confirming uORF-mediated translational potentiation of the cognate main ORF. This study establishes uORF engineering as a robust platform for predictable metabolic engineering in S. miltiorrhiza plants. Future applications could expand this strategy to uORFs of rate-limiting enzymes or transcriptional regulators, enabling multidimensional optimization of high-value metabolites in medicinal species.<\/jats:p>","DOI":"10.1093\/hr\/uhaf249","type":"journal-article","created":{"date-parts":[[2025,9,20]],"date-time":"2025-09-20T05:31:26Z","timestamp":1758346286000},"source":"Crossref","is-referenced-by-count":9,"title":["CRISPR\/Cas9-mediated uORF engineering enhances tanshinone biosynthesis in\n                    <i>Salvia miltiorrhiza<\/i>"],"prefix":"10.1093","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1157-1565","authenticated-orcid":false,"given":"Jin","family":"Shao","sequence":"first","affiliation":[{"name":"Frontiers Science Center for Transformative Molecules, Joint International Research Laboratory of Metabolic and Developmental Sciences, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University , Shanghai 200240 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3575-282X","authenticated-orcid":false,"given":"Bowen","family":"Peng","sequence":"additional","affiliation":[{"name":"Frontiers Science Center for Transformative Molecules, Joint International Research Laboratory of Metabolic and Developmental Sciences, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University , Shanghai 200240 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Han","family":"Zheng","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences , Beijing 100700 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5194-0391","authenticated-orcid":false,"given":"Ling","family":"Li","sequence":"additional","affiliation":[{"name":"Frontiers Science Center for Transformative Molecules, Joint International Research Laboratory of Metabolic and Developmental Sciences, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University , Shanghai 200240 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dalu","family":"Li","sequence":"additional","affiliation":[{"name":"Shanghai Agricultural Technology Extension Service Center , Shanghai 201103 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8172-2448","authenticated-orcid":false,"given":"Xinyi","family":"Hu","sequence":"additional","affiliation":[{"name":"Frontiers Science Center for Transformative Molecules, Joint International Research Laboratory of Metabolic and Developmental Sciences, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University , Shanghai 200240 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luqi","family":"Huang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences , Beijing 100700 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8748-9624","authenticated-orcid":false,"given":"Kexuan","family":"Tang","sequence":"additional","affiliation":[{"name":"Frontiers Science Center for Transformative Molecules, Joint International Research Laboratory of Metabolic and Developmental Sciences, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University , Shanghai 200240 ,","place":["China"]},{"name":"Yazhouwan National Laboratory , Sanya, Hainan 572024 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2025,9,17]]},"reference":[{"key":"2026013005570649800_ref1","first-page":"163","article-title":"Salvia miltiorrhiza: traditional medicinal uses, chemistry, and pharmacology","volume":"13","author":"Su","year":"2015","journal-title":"Chin J Nat 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