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Dissection of miRNA Pathways Using Arabidopsis Mesophyll Protoplasts

2015-02-02Journal article Open access
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dc.contributor.authorMartinho, Cláudia
dc.contributor.authorConfraria, Ana
dc.contributor.authorElias, Carlos Alexandre
dc.contributor.authorCrozet, Pierre
dc.contributor.authorRubio-Somoza, Ignacio
dc.contributor.authorWeigel, Detlef
dc.contributor.authorBaena-González, Elena
dc.date.accessioned2016-06-07T14:38:11Z
dc.date.available2016-06-07T14:38:11Z
dc.date.issued2015-02-02
dc.description.abstractMicroRNAs (miRNAs) control gene expression mostly post-transcriptionally by guiding transcript cleavage and/or translational repression of complementary mRNA targets, thereby regulating developmental processes and stress responses. Despite the remarkable expansion of the field, the mechanisms underlying miRNA activity are not fully understood. In this article, we describe a transient expression system in Arabidopsis mesophyll protoplasts, which is highly amenable for the dissection of miRNA pathways. We show that by transiently overexpressing primary miRNAs and target mimics, we can manipulate miRNA levels and consequently impact on their targets. Furthermore, we developed a set of luciferase-based sensors for quantifying miRNA activity that respond specifically to both endogenous and overexpressed miRNAs and target mimics. We demonstrate that these miRNA sensors can be used to test the impact of putative components of the miRNA pathway on miRNA activity, as well as the impact of specific mutations, by either overexpression or the use of protoplasts from the corresponding mutants. We further show that our miRNA sensors can be used for investigating the effect of chemicals on miRNA activity. Our cell-based transient expression system is fast and easy to set up, and generates quantitative results, being a powerful tool for assaying miRNA activity in vivo.pt_PT
dc.description.sponsorshipFundação para a Ciência e Tecnologia fellowships: (SFRH/BD/33563/2008, SFRH/BPD/47280/2008, SFRH/BPD/79255/2011) and grant: (PTCD/BIA-BCM/107924/2008); EMBO fellowship & EMBO Installation program; Deutsche Forschungsgemeinschaft grant: (SPP1530); Max Planck Society grant.pt_PT
dc.identifier.citationCláudia Martinho, Ana Confraria, Carlos Alexandre Elias, Pierre Crozet, Ignacio Rubio-Somoza, Detlef Weigel, Elena Baena-González, Dissection of miRNA Pathways Using Arabidopsis Mesophyll Protoplasts, Molecular Plant, Volume 8, Issue 2, 2 February 2015, Pages 261-275, ISSN 1674-2052, http://dx.doi.org/10.1016/j.molp.2014.10.003.pt_PT
dc.identifier.doi10.1016/j.molp.2014.10.003pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.7/624
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherCell Presspt_PT
dc.relation.publisherversionhttp://www.sciencedirect.com/science/article/pii/S1674205214000185pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectArabidopsispt_PT
dc.subjectGene Expression Regulation, Plantpt_PT
dc.subjectMicroRNAspt_PT
dc.subjectProtoplastspt_PT
dc.titleDissection of miRNA Pathways Using Arabidopsis Mesophyll Protoplastspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage275pt_PT
oaire.citation.issue2pt_PT
oaire.citation.startPage261pt_PT
oaire.citation.titleMolecular Plantpt_PT
oaire.citation.volume8pt_PT
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT

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