Custom-made design of metabolite composition in N. benthamiana leaves using CRISPR activators
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Título
Custom-made design of metabolite composition in N. benthamiana leaves using CRISPR activatorsAutoría
Fecha de publicación
2022-05-05Editor
WileyISSN
1467-7644; 1467-7652Cita bibliográfica
Selma, S., Sanmartín, N., Espinosa-Ruiz, A., Gianoglio, S., Lopez-Gresa, M. P., Vázquez-Vilar, M., Flors, V., Granell, A. and Orzaez, D. (2022) Custom-made design of metabolite composition in N. benthamiana leaves using CRISPR activators. Plant Biotechnol. J.Tipo de documento
info:eu-repo/semantics/articleVersión
info:eu-repo/semantics/publishedVersionPalabras clave / Materias
Resumen
Transcriptional regulators based on CRISPR architecture expand our ability to reprogramme endogenous gene expression in plants. One of their potential applications is the customization of plant metabolome through the ... [+]
Transcriptional regulators based on CRISPR architecture expand our ability to reprogramme endogenous gene expression in plants. One of their potential applications is the customization of plant metabolome through the activation of selected enzymes in a given metabolic pathway. Using the previously described multiplexable CRISPR activator dCasEV2.1, we assayed the selective enrichment in Nicotiana benthamiana leaves of four different flavonoids, namely, naringenin, eriodictyol, kaempferol, and quercetin. After careful selection of target genes and guide RNAs combinations, we created successful activation programmes for each of the four metabolites, each programme activating between three and seven genes, and with individual gene activation levels ranging from 4- to 1500-fold. Metabolic analysis of the flavonoid profiles of each multigene activation programme showed a sharp and selective enrichment of the intended metabolites and their glycosylated derivatives. Remarkably, principal component analysis of untargeted metabolic profiles clearly separated samples according to their activation treatment, and hierarchical clustering separated the samples into five groups, corresponding to the expected four highly enriched metabolite groups, plus an un-activated control. These results demonstrate that dCasEV2.1 is a powerful tool for re-routing metabolic fluxes towards the accumulation of metabolites of interest, opening the door for the custom-made design of metabolic contents in plants. [-]
Publicado en
Plant Biotechnology Journal 20 (2022)Entidad financiadora
Ministerio de Ciencia, Innovación y Universidades | Ministerio de Economía y Competitividad
Código del proyecto o subvención
PID2019-108203RB-10 | BIO2016- 78601-R
Título del proyecto o subvención
Plan Nacional I + D
Derechos de acceso
info:eu-repo/semantics/openAccess
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