{"id":100162,"date":"2010-09-04T00:00:00","date_gmt":"2010-09-04T00:00:00","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/generacion-de-ratones-transgenicos-con-expresion-especa%c2%adfica-de-recombinasa-cre-en-megacariocitos\/"},"modified":"2010-09-04T00:00:00","modified_gmt":"2010-09-04T00:00:00","slug":"generacion-de-ratones-transgenicos-con-expresion-especa%c2%adfica-de-recombinasa-cre-en-megacariocitos","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/generacion-de-ratones-transgenicos-con-expresion-especa%c2%adfica-de-recombinasa-cre-en-megacariocitos\/","title":{"rendered":"Generaci\u00f3n de ratones transg\u00e9nicos con expresi\u00f3n espec\u00edfica de recombinasa cre en megacariocitos."},"content":{"rendered":"<h2>Tesis doctoral de <strong> Adam Nowakowski <\/strong><\/h2>\n<p>Para poder estudiar los efectos de la anulaci\u00f3n de expresi\u00f3n de los genes cuyos productos proteicos son cruciales durante el desarrollo, se generan los ratones \u00c2\u00bfknock out\u00c2\u00bf condicionales. En estos animales, se induce la anulaci\u00f3n de expresi\u00f3n del gen de inter\u00e9s en el animal adulto, lo que permite un desarrollo embrionario normal. Para generar este tipo de animales, se emplea el sistema de recombinaci\u00f3n cre\/loxp. el sistema cre-loxp es la estrategia m\u00e1s utilizada en la mutag\u00e9nesis dirigida, espec\u00ed fica de tejido. La colocaci\u00f3n y orientaci\u00f3n de los sitios loxp (que se introducen en la secuencia de adn a recombinar) determinar\u00e1n la secuencia gen\u00f3mica a escindir, mientras que la disponibilidad, en el tiempo y\/o el espacio, de la recombinasa cre, que se obtiene a partir de un transg\u00e9n dictar\u00e1 cuando y d\u00f3nde se produce la recombinaci\u00f3n y, por tanto, el \u00c2\u00bfknock-out\u00c2\u00bf. Los sitios loxp se disponen en parejas flanqueando el segmento de adn a eliminar. Cuando cre se expresa, la recombinasa cre se une  a los sitios loxp, los corta por la mitad y despu\u00e9s une las dos mitades restantes tras haber eliminado el adn situado entre ambos. el objetivo principal de nuestro trabajo ha sido la generaci\u00f3n de ratones transg\u00e9nicos con expresi\u00f3n espec\u00edfica de cre  en megacariocitos y plaquetas y su verificaci\u00f3n funcional. generamos ratones transg\u00e9nicos mediante microinyecci\u00f3n de nuestra construcci\u00f3n de adn en pron\u00facleos de oocitos fecundados. Conseguimos generar un total de 4 l\u00edneas de animales con transmisi\u00f3 n del transg\u00e9n por v\u00eda germinal. la intensidad de la expresi\u00f3n del transg\u00e9n cre la verificamos mediante generaci\u00f3n de ratones doble transg\u00e9nicos m-alfaiib-cre \/ rosa26-lacz. En estos animales la acci\u00f3n de la recombinasa cre gener\u00f3 cantidad suficiente  de la beta-galactosidasa como para desarrollar color de x-gal espec\u00edficamente en megacariocitos y plaquetas. \tnuestros resultados abren la posibilidad de explorar numerosas etapas de control, tanto metab\u00f3licas como del desarrollo, actuando sobre gen es \u00c2\u00bffloxed\u00c2\u00bf de importancia reguladora en megacariocitos. Dada la viabilidad as\u00ed como la ausencia de alteraciones funcionales detectables, los ratones transg\u00e9nicos m-alfaiib-cre, ofrecen una herramienta de potencial valor biol\u00f3gico y de aplicabilidad inmediata.<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Generaci\u00f3n de ratones transg\u00e9nicos con expresi\u00f3n espec\u00edfica de recombinasa cre en megacariocitos.<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Generaci\u00f3n de ratones transg\u00e9nicos con expresi\u00f3n espec\u00edfica de recombinasa cre en megacariocitos. <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Adam Nowakowski <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Complutense de Madrid<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 09\/04\/2010<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3>Direcci\u00f3n y tribunal<\/h3>\n<ul>\n<li><strong>Director de la tesis<\/strong>\n<ul>\n<li>Roberto Parrilla S\u00e1nchez<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: Mar\u00eda teresa P\u00f3rtoles p\u00e9rez <\/li>\n<li>Carlos Guti\u00e9rrez merino (vocal)<\/li>\n<li>carmelo Bernabeu quirante (vocal)<\/li>\n<li>matilde S\u00e1nchez ayuso (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Adam Nowakowski Para poder estudiar los efectos de la anulaci\u00f3n de expresi\u00f3n de los genes cuyos productos [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center 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