dc.contributor.author |
Vázquez-Vilar, Marta
|
es_ES |
dc.contributor.author |
Bernabé-Orts, Joan Miquel
|
es_ES |
dc.contributor.author |
Fernández Del Carmen, María Asunción
|
es_ES |
dc.contributor.author |
Ziarsolo Areitioaurtena, Pello
|
es_ES |
dc.contributor.author |
Blanca Postigo, José Miguel
|
es_ES |
dc.contributor.author |
Granell Richart, Antonio
|
es_ES |
dc.contributor.author |
Orzáez Calatayud, Diego Vicente
|
es_ES |
dc.date.accessioned |
2018-02-02T10:07:04Z |
|
dc.date.available |
2018-02-02T10:07:04Z |
|
dc.date.issued |
2016 |
es_ES |
dc.identifier.issn |
1746-4811 |
es_ES |
dc.identifier.uri |
http://hdl.handle.net/10251/96423 |
|
dc.description.abstract |
[EN] Background: The efficiency, versatility and multiplexing capacity of RNA-guided genome engineering using the
CRISPR/Cas9 technology enables a variety of applications in plants, ranging from gene editing to the construction
of transcriptional gene circuits, many of which depend on the technical ability to compose and transfer complex
synthetic instructions into the plant cell. The engineering principles of standardization and modularity applied to DNA
cloning are impacting plant genetic engineering, by increasing multigene assembly efficiency and by fostering the
exchange of well-defined physical DNA parts with precise functional information.
Results: Here we describe the adaptation of the RNA-guided Cas9 system to GoldenBraid (GB), a modular DNA con¿
struction framework being increasingly used in Plant Synthetic Biology. In this work, the genetic elements required
for CRISPRs-based editing and transcriptional regulation were adapted to GB, and a workflow for gRNAs construction
was designed and optimized. New software tools specific for CRISPRs assembly were created and incorporated to the
public GB resources site.
Conclusions: The functionality and the efficiency of gRNA¿Cas9 GB tools were demonstrated in Nicotiana benthamiana
using transient expression assays both for gene targeted mutations and for transcriptional regulation. The
availability of gRNA¿Cas9 GB toolbox will facilitate the application of CRISPR/Cas9 technology to plant genome
engineering |
es_ES |
dc.description.sponsorship |
This work has been funded by Grant BIO2013-42193-R from Plan Nacional I + D of the Spanish Ministry of Economy and Competitiveness. Vazquez-Vilar M. is a recipient of a Junta de Ampliacion de Estudios fellowship. Bernabe-Orts J.M. is a recipient of a FPI fellowship. We want to thank Nicola J. Patron and Mark Youles for kindly providing humanCas9 and U6-26 clones. We also want to thank Eugenio Gomez for providing Arabidopsis thaliana genomic DNA and Concha Domingo for providing rice genomic DNA. We also want to thank the COST Action FA1006 for the support in the development of the software tools. |
es_ES |
dc.language |
Inglés |
es_ES |
dc.publisher |
Springer (Biomed Central Ltd.) |
es_ES |
dc.relation.ispartof |
Plant Methods |
es_ES |
dc.rights |
Reconocimiento (by) |
es_ES |
dc.subject |
Plant gene editing |
es_ES |
dc.subject |
Plant gene activation |
es_ES |
dc.subject |
Plant gene repression |
es_ES |
dc.subject |
CRISPR/Cas9 |
es_ES |
dc.subject |
gRNAs |
es_ES |
dc.subject |
Multigenic assemblies |
es_ES |
dc.subject |
GoldenBraid |
es_ES |
dc.subject |
Luciferase/renilla assay |
es_ES |
dc.subject.classification |
GENETICA |
es_ES |
dc.subject.classification |
BIOQUIMICA Y BIOLOGIA MOLECULAR |
es_ES |
dc.title |
A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard |
es_ES |
dc.type |
Artículo |
es_ES |
dc.identifier.doi |
10.1186/s13007-016-0101-2 |
es_ES |
dc.relation.projectID |
info:eu-repo/grantAgreement/MINECO//BIO2013-42193-R/ES/GREEN SWITCHES: DISEÑO DE CIRCUITOS GENETICOS ARTIFICIALES PARA LA PRODUCCION DE PROTEINAS RECOMBINANTES Y EL ENRIQUECIMIENTO NUTRICIONAL DE PLANTAS SOLANACEAS/ |
es_ES |
dc.rights.accessRights |
Abierto |
es_ES |
dc.contributor.affiliation |
Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes |
es_ES |
dc.contributor.affiliation |
Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia |
es_ES |
dc.contributor.affiliation |
Universitat Politècnica de València. Instituto Universitario de Conservación y Mejora de la Agrodiversidad Valenciana - Institut Universitari de Conservació i Millora de l'Agrodiversitat Valenciana |
es_ES |
dc.description.bibliographicCitation |
Vázquez-Vilar, M.; Bernabé-Orts, JM.; Fernández Del Carmen, MA.; Ziarsolo Areitioaurtena, P.; Blanca Postigo, JM.; Granell Richart, A.; Orzáez Calatayud, DV. (2016). A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard. Plant Methods. 12. https://doi.org/10.1186/s13007-016-0101-2 |
es_ES |
dc.description.accrualMethod |
S |
es_ES |
dc.relation.publisherversion |
http://doi.org/10.1186/s13007-016-0101-2 |
es_ES |
dc.type.version |
info:eu-repo/semantics/publishedVersion |
es_ES |
dc.description.volume |
12 |
es_ES |
dc.subject.asignatura |
Aplicaciones de la biotecnología al diseño de nuevos carácteres y productos 32722 / W - Programa de doctorado en biotecnología 2071 |
es_ES |
dc.subject.asignatura |
Aplicaciones de la biotecnología al diseño de nuevos carácteres y productos 32722 / X - Máster universitario en biotecnología molecular y celular de plantas 2172 |
es_ES |
dc.identifier.pmid |
26839579 |
|
dc.identifier.pmcid |
PMC4736081 |
|
dc.relation.pasarela |
S\306679 |
es_ES |
dc.contributor.funder |
Ministerio de Economía y Competitividad |
es_ES |
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