Análisis de las interacciones microRNAs-lncRNAs en el proepicardio de pollo
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2019-06-14
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Jaén: Universidad de Jaén
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[ES] Los nuevos avances en el estudio del genoma han demostrado el importante papel del RNA no codificante en muchos procesos biológicos. El desarrollo cardiaco es un proceso determinante para la supervivencia adulta, su complejidad hace suponer que muchas cardiopatías tengan su origen en la cardiogénesis. Recientes estudios en la UJA han demostrado la expresión de microRNAs y lncRNAs implicados en el destino del linaje celular cardiaco, en proepicardio. En el presente estudio, utilizamos proepicardios (PE) de pollo en HH17 para sobre-expresar distintos miRNAs. Analizamos en este órgano, la posible regulación de forma diferencial de la expresión de algunos lncRNAs por parte de los miRNAs. Los PE fueron transfectados con premiRNAs mediante lipoesferas para posteriormente extraer la totalidad del RNA de la muestra. Tras la lectura de purificación, este RNA se retrotranscribió para la realización de dos q-PCRs. En primer lugar, para observar la correcta sobre-expresión de miRNAs. En segundo lugar, con primers de lncRNAs para localizarlos y evaluar su expresión con la influencia de los miRNAs sobre- expresados. Por último, se muestra la búsqueda de ORFs en las secuencias polipeptídicas de los lncRNAs estudiados, en concreto secuencias de importancia en la cardiogénesis de aves.
[EN] New advances in the study of genomics have demonstrated the important role of non-coding RNA in a lot of biological processes. Cardiac development is a determining process for adult survival, its complexity suggests that many heart desseas have their origin in cardiac development. Recent studies from The University of Jaén have shown the expression of microRNAs and lncRNAs involved in the fate of the cardiac cell lineage (specifically in proepicardium). In the recent study, we used chicken proepicardium (PE) in HH17 to over-express diferent miRNAs (miR-23, miR-27, miR-195 y miR-223). We analyze in this organ, the posible differential regulation of some of the lncRNAs (Wt1_76127, Bmp4_53170 and Fgf8_57126) by the miRNAs. The PE were transfected with premiRNAs by means of lipospheres to later extract all the RNA from the sample. After reading the purification, this RNA was retrotranscribed to do q-PCR twice. First, to observe the cocrrect over-expressed miRNAs. Secondly, with primers of lncRNAs to localize them and evaluate their expression with the influence of the over-expressed miRNAs. Finaly, it shows the search for ORFs in the polypeptide secuences of the lncRNAs studied, in particular important secuences in heart development of birds.
[EN] New advances in the study of genomics have demonstrated the important role of non-coding RNA in a lot of biological processes. Cardiac development is a determining process for adult survival, its complexity suggests that many heart desseas have their origin in cardiac development. Recent studies from The University of Jaén have shown the expression of microRNAs and lncRNAs involved in the fate of the cardiac cell lineage (specifically in proepicardium). In the recent study, we used chicken proepicardium (PE) in HH17 to over-express diferent miRNAs (miR-23, miR-27, miR-195 y miR-223). We analyze in this organ, the posible differential regulation of some of the lncRNAs (Wt1_76127, Bmp4_53170 and Fgf8_57126) by the miRNAs. The PE were transfected with premiRNAs by means of lipospheres to later extract all the RNA from the sample. After reading the purification, this RNA was retrotranscribed to do q-PCR twice. First, to observe the cocrrect over-expressed miRNAs. Secondly, with primers of lncRNAs to localize them and evaluate their expression with the influence of the over-expressed miRNAs. Finaly, it shows the search for ORFs in the polypeptide secuences of the lncRNAs studied, in particular important secuences in heart development of birds.