A novel RT-QPCR-based assay for the relative quantification of residue specific m6A RNA methylation
dc.contributor.author | Castellanos Rubio, Ainara | |
dc.contributor.author | Santín Gómez, Izortze | |
dc.contributor.author | Olazagoitia Garmendia, Ane | |
dc.contributor.author | Romero Garmendia, Irati | |
dc.contributor.author | Jauregi Miguel, Amaia | |
dc.contributor.author | Legarda Tamara, María | |
dc.contributor.author | Bilbao Catalá, José Ramón | |
dc.date.accessioned | 2019-04-04T18:00:28Z | |
dc.date.available | 2019-04-04T18:00:28Z | |
dc.date.issued | 2019-12 | |
dc.identifier.citation | Scientific Reports (9)1: (2019) // Article ID 4220 | es_ES |
dc.identifier.issn | 2045-2322 | |
dc.identifier.uri | http://hdl.handle.net/10810/32330 | |
dc.description.abstract | N6-methyladenosine (m6A) is the most common and abundant RNA modification. Recent studies have shown its importance in the regulation of several biological processes, including the immune response, and different approaches have been developed in order to map and quantify m6A marks. However, site specific detection of m6A methylation has been technically challenging, and existing protocols are long and tedious and often involve next-generation sequencing. Here, we describe a simple RT-QPCR based approach for the relative quantification of candidate m6A regions that takes advantage of the diminished capacity of BstI enzyme to retrotranscribe m6A residues. Using this technique, we have been able to confirm the recently described m6A methylation in the 3′UTR of SOCS1 and SOCS3 transcripts. Moreover, using the method presented here, we have also observed alterations in the relative levels of m6A in specific motifs of SOCS genes in celiac disease patients and in pancreatic β-cells exposed to inflammatory stimuli. | es_ES |
dc.description.sponsorship | J.R.B. is funded by Project ISCIII-PI16/00258 and co-funded by the European Union ERDF/ESF “A way to make Europe”. I.S. is funded by a research project grant 2015111068 of the Basque Department of Health. A.C.R. was funded by a Juan de la Cierva reincorporation and an Ikerbasque fellowships and a research project grant from Asociación Celiacos Madrid. I.R.G., A.O.G. and A.J.M. are supported by predoctoral fellowship grants from the UPV/EHU and the Basque Department of Education. | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | Nature Publishing Group | es_ES |
dc.rights | info:eu-repo/semantics/openAccess | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/es/ | * |
dc.title | A novel RT-QPCR-based assay for the relative quantification of residue specific m6A RNA methylation | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.rights.holder | © The Author(s) 2019.This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. | es_ES |
dc.rights.holder | Atribución 3.0 España | * |
dc.relation.publisherversion | https://www.nature.com/articles/s41598-019-40018-6 | es_ES |
dc.identifier.doi | 10.1038/s41598-019-40018-6 | |
dc.departamentoes | Bioquímica y biología molecular | es_ES |
dc.departamentoes | Genética, antropología física y fisiología animal | es_ES |
dc.departamentoeu | Biokimika eta biologia molekularra | es_ES |
dc.departamentoeu | Genetika,antropologia fisikoa eta animalien fisiologia | es_ES |
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Except where otherwise noted, this item's license is described as © The Author(s) 2019.This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.