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This study investigates the bioengineering potential of the marine diatom Phaeodactylum tricornutum through the introduction of cannabis genes, specifically, tetraketide synthase (TKS), and olivetolic acid cyclase (OAC), for the production of the cannabinoid precursor, olivetolic acid (OA). P. tricornutum is a promising biotechnological platform due to its fast growth rate, amenability to genetic manipulation, and ability to produce valuable compounds. Through genetic engineering techniques, we successfully integrated the cannabis genes TKS and OAC into the diatom. P. tricornutum transconjugants expressing these genes showed the production of the recombinant TKS and OAC enzymes, detected via Western blot analysis, and the production of cannabinoids precursor (OA) detected using the HPLC\/UV spectrum when compared to the wild-type strain. Quantitative analysis revealed significant olivetolic acid accumulation (0.6\u20132.6 mg\/L), demonstrating the successful integration and functionality of the heterologous genes. Furthermore, the introduction of TKS and OAC genes led to the synthesis of novel molecules, potentially expanding the repertoire of bioactive compounds accessible through diatom-based biotechnology. This study demonstrates the successful bioengineering of P. tricornutum with cannabis genes, enabling the production of OA as a precursor for cannabinoid production and the synthesis of novel molecules with potential pharmaceutical applications.<\/jats:p>","DOI":"10.3390\/ijms242316624","type":"journal-article","created":{"date-parts":[[2023,11,22]],"date-time":"2023-11-22T08:47:43Z","timestamp":1700642863000},"page":"16624","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":27,"title":["Bioengineering of the Marine Diatom Phaeodactylum tricornutum with Cannabis Genes Enables the Production of the Cannabinoid Precursor, Olivetolic Acid"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0428-5552","authenticated-orcid":false,"given":"Fatima","family":"Awwad","sequence":"first","affiliation":[{"name":"Department of Chemistry, Biochemistry and Physics, Universit\u00e9 du Qu\u00e9bec \u00e0 Trois-Rivi\u00e8res, 3351 Boulevard des Forges, Trois-Riviere, QC G9A 5H7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8393-3241","authenticated-orcid":false,"given":"Elisa Ines","family":"Fantino","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Biochemistry and Physics, Universit\u00e9 du Qu\u00e9bec \u00e0 Trois-Rivi\u00e8res, 3351 Boulevard des Forges, Trois-Riviere, QC G9A 5H7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marianne","family":"H\u00e9neault","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Biochemistry and Physics, Universit\u00e9 du Qu\u00e9bec \u00e0 Trois-Rivi\u00e8res, 3351 Boulevard des Forges, Trois-Riviere, QC G9A 5H7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1923-2974","authenticated-orcid":false,"given":"Aracely Maribel","family":"Diaz-Garza","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Biochemistry and Physics, Universit\u00e9 du Qu\u00e9bec \u00e0 Trois-Rivi\u00e8res, 3351 Boulevard des Forges, Trois-Riviere, QC G9A 5H7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6235-6465","authenticated-orcid":false,"given":"Natacha","family":"Merindol","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Biochemistry and Physics, Universit\u00e9 du Qu\u00e9bec \u00e0 Trois-Rivi\u00e8res, 3351 Boulevard des Forges, Trois-Riviere, QC G9A 5H7, Canada"},{"name":"Groupe de Recherche en Biologie V\u00e9g\u00e9tale, Universit\u00e9 du Qu\u00e9bec \u00e0 Trois-Rivi\u00e8res, Trois-Riviere, QC G9A 5H7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre","family":"Custeau","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Biochemistry and Physics, 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