Caracterización de la expresión de lncRNAs durante el desarrollo cardiovascular.
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2018-03-13
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Jaén: Universidad de Jaén
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[ES] Una gran parte del genoma de ratón no contiene genes codificantes, por lo que no se traducen a proteínas, pero sí que se transcriben de manera activa dando lugar a diferentes tipos de ARNs. Aquí encontramos los long-non-codings RNAs (lncRNAs) que son secuencias no codificantes de un tamaño considerable (más de 200 nucleótidos de longitud) del genoma de ratón. El estudio de este genoma no codificante es relativamente reciente, ya que no se había considerado relevante hasta que se ha visto que estos lncRNAs pueden contribuir a la regulación de ciertos procesos celulares, y con ello, que pueden afectar también en algunas enfermedades. Tras numerosos estudios se ha descubierto que la expresión de los lncRNAs se regula durante el desarrollo y que, además, esta expresión es tejido específico.
En nuestro laboratorio hemos analizado el perfil de expresión de diferentes lncRNAs cardiovasculares, en concreto para Braveheart, Carmen, Fendrr, H19 y Miat, mostrando diferencias entre las distintas cámaras cardíacas. Para ello, queremos obtener sondas específicas. Por tanto, se han diseñado primers para los lncRNAs, para amplificarlos mediante PCRs, y clonarlos en vectores plasmídicos, que permitían la inserción del lncRNA en cuestión. En concreto, hemos obtenido 4 sondas específicas para Braveheart, Fendrr, H19 y Miat, de las cuales hemos verificado la secuencia y la dirección del inserto.
Nuestros resultados permiten obtener sondas para realizar la hibridación in situ y poder determinar la localización histológica y el nivel de expresión de estos lncRNAs en diferentes etapas del desarrollo cardíaco en ratón.
[EN] A large part of the mouse genome does not contain coding genes, so they are not translated into proteins, but they are actively transcribed giving rise to different types of RNAs. Here we find the long-non-coding RNAs (lncRNAs) which are non-coding sequences of considerable size (more than 200 nucleotides in length) from the mouse genome. The study of this non-coding geno me is relatively recent, since it had not been considered relevant until it has been seen that these lncRNAs can contribute to the regulation of certain cellular processes, and with that, can also affect sorne diseases. After numerous studies, it has been discovered that the expression of lncRNAs is regulated during development and that, in addition, this expression is a specific tissue. In our laboratory, we have analyzed the expression profile of different cardiovascular lncRNAs, specifically for Braveheart, Carmen, Fendrr, H19 and Miat, showing differences between the different cardiac chambers. For that, we want to obtain specific probes. Therefore, primers have been designed for lncRNAs, amplified by PCRs, and cloned into plasmid vectors, which allowed the insertion of the lncRNA in question. In particular, we have obtained 4 specific probes for Braveheart, Fendrr, H19 and Miat, from which we have verified the sequence and direction of the insert. Our results allow to obtain probes to perform the hybridization in situ and to be able to determine the histological location and the level of expression of these lncRNAs in different stages of the cardiac development in mice.
[EN] A large part of the mouse genome does not contain coding genes, so they are not translated into proteins, but they are actively transcribed giving rise to different types of RNAs. Here we find the long-non-coding RNAs (lncRNAs) which are non-coding sequences of considerable size (more than 200 nucleotides in length) from the mouse genome. The study of this non-coding geno me is relatively recent, since it had not been considered relevant until it has been seen that these lncRNAs can contribute to the regulation of certain cellular processes, and with that, can also affect sorne diseases. After numerous studies, it has been discovered that the expression of lncRNAs is regulated during development and that, in addition, this expression is a specific tissue. In our laboratory, we have analyzed the expression profile of different cardiovascular lncRNAs, specifically for Braveheart, Carmen, Fendrr, H19 and Miat, showing differences between the different cardiac chambers. For that, we want to obtain specific probes. Therefore, primers have been designed for lncRNAs, amplified by PCRs, and cloned into plasmid vectors, which allowed the insertion of the lncRNA in question. In particular, we have obtained 4 specific probes for Braveheart, Fendrr, H19 and Miat, from which we have verified the sequence and direction of the insert. Our results allow to obtain probes to perform the hybridization in situ and to be able to determine the histological location and the level of expression of these lncRNAs in different stages of the cardiac development in mice.