Transfecting taxus x media protoplasts to study transcription factors BIS2 and TSAR2 as activators of taxane-related genes

Taxane diterpenes are secondary metabolites with an important pharmacological role in the treatment of cancer. Taxus spp. biofactories have been used for taxane production, but the lack of knowledge about the taxane biosynthetic pathway and its molecular regulation hinders their optimal function. Th...

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Detalles Bibliográficos
Autores: Sánchez Muñoz, Raúl|||0000-0003-0235-2985, Almagro, Lorena, Cusido, Rosa M, Bonfill, Mercedes, Palazon, Javier, Moyano, Elisabeth
Tipo de recurso: artículo
Fecha de publicación:2020
País:España
Institución:Universitat Politècnica de Catalunya (UPC)
Repositorio:UPCommons. Portal del coneixement obert de la UPC
Idioma:inglés
OAI Identifier:oai:upcommons.upc.edu:2117/447100
Acceso en línea:https://hdl.handle.net/2117/447100
https://dx.doi.org/10.1093/pcp/pcz225
Access Level:acceso abierto
Palabra clave:Taxus¿x¿media
Metabolic engineering
Protoplasts
Taxanes
Transcription factors
Transient transfection.
Àrees temàtiques de la UPC::Enginyeria agroalimentària::Agricultura::Biotecnologia i millora genètica vegetal
Descripción
Sumario:Taxane diterpenes are secondary metabolites with an important pharmacological role in the treatment of cancer. Taxus spp. biofactories have been used for taxane production, but the lack of knowledge about the taxane biosynthetic pathway and its molecular regulation hinders their optimal function. The difficulties in introducing foreign genes in Taxus spp. genomes hinder the study of the molecular mechanisms involved in taxane production, and a new approach is required to overcome them. In this study, a reliable, simple and fast method to obtain Taxus¿×¿media protoplasts was developed, allowing their manipulation in downstream assays for the study of physiological changes in Taxus spp. cells. Using this method, Taxus protoplasts were transiently transfected for the first time, corroborating their suitability for transfection assays and the study of specific physiological responses. The two assayed transcription factors (BIS2 and TSAR2) had a positive effect on the expression of several taxane-related genes, suggesting their potential use for the improvement of taxane yields. Furthermore, the results indicate that the developed method is suitable for obtaining T.¿×¿media protoplasts for transfection with the aim of unraveling regulatory mechanisms in taxane production.