Clonación de elementos reguladores de lncRNAs implicados en el desarrollo cardiovascular y análisis funcionales
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2021-07-06
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Jaén: Universidad de Jaén
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[ES] En un estudio anterior, se identificaron tres nuevos lncRNAs que se encuentran en explantes de PE/ST y que son capaces de regular una serie de microRNAs que actúan sobre el desarrollo del PE y del epicardio. Pero aún no se conoce la regulación de estos lncRNAs. Con este trabajo se pretende identificar y analizar el posible sistema de regulación transcripcional de estos lncRNAs. Para ello, analizamos el nivel de conservación en otras especies, diseñamos primers para los promotores de estos lncRNAs, posteriormente se amplificó y clonó en pGemT y pGluc, y estudiamos su actividad transcripcional en células de 3T3 cuando se le adicionan unos factores de transcripción. Con este proyecto se confirmó la no conservación entre especies de los promotores. Se consiguió amplificar y clonar la región promotora de la isoforma 2 de Wt1_76127 y se estudió el nivel de transactivación que producen estos factores del gen de la luciferasa.
[EN] In a previous study, three new lncRNAs were identified that are found in PE / ST explants and they are capable of regulating a series of microRNAs that act on the development of the PE and the epicardium. But it is not yet known how these lncRNAs may be regulated. This proyect aims to identify and analyze the possible transcriptional regulation system of these lncRNAs. To do this, we analyzed their level of conservation in other species, we designed primers for the promoters of these lncRNAs, later it was amplified and cloned in pGemT and pGluc, and we studied their transcriptional activity in 3T3 cells when some transcription factors were added. With this project, the non-conservation between species of the promotors of these lncRNAs was confirmed. The promotor of isoform 2 of Wt1_76127 was amplified and cloned, and we studied the level of transactivation produced by these factors on the luciferase gene.
[EN] In a previous study, three new lncRNAs were identified that are found in PE / ST explants and they are capable of regulating a series of microRNAs that act on the development of the PE and the epicardium. But it is not yet known how these lncRNAs may be regulated. This proyect aims to identify and analyze the possible transcriptional regulation system of these lncRNAs. To do this, we analyzed their level of conservation in other species, we designed primers for the promoters of these lncRNAs, later it was amplified and cloned in pGemT and pGluc, and we studied their transcriptional activity in 3T3 cells when some transcription factors were added. With this project, the non-conservation between species of the promotors of these lncRNAs was confirmed. The promotor of isoform 2 of Wt1_76127 was amplified and cloned, and we studied the level of transactivation produced by these factors on the luciferase gene.