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Reconstrucción molecular de la Bella Durmiente, un transposón similar al Tc1 de los peces, y su transposición en

Z Ivics¹, P B Hackett, R H Plasterk

¹Department of Genetics and Cell Biology, University of Minnesota, St. Paul 55108-1095, USA.

|December 9, 1997

Resumen

Los investigadores diseñaron el transposón de la Bella Durmiente, un sistema sintético de ADN-transposón, mediante la reconstrucción de un antiguo elemento de pez. Este sistema activo permite una transposición precisa de corte y pega para la transformación genética y la mutagénesis por inserción en varias células de vertebrados.

Área de la Ciencia:

Genética La genética.
Biología Molecular Biología Molecular
Biología evolutiva Biología evolutiva.

Sus antecedentes:

Las transposones Tc1/mariner en los peces suelen estar inactivas debido a mutaciones acumuladas.
Los antiguos elementos del transposón pueden haberse dispersado a través de la transferencia horizontal de genes entre especies de peces.

Objetivo del estudio:

Para reconstruir un sistema funcional de ADN-transposón a partir de elementos de peces inactivos.
Diseñar un transposón activo para la manipulación genética en vertebrados.

Principales métodos:

Utilizó datos filogenéticos moleculares para diseñar un transposón sintético, la Bella Durmiente.
Diseñó un gen de la transposasa de consenso mediante la eliminación de mutaciones inactivadoras.
Evaluó la actividad del ADN-transposón de la Bella Durmiente en peces, ratones y células humanas.

Principales resultados:

La transposasa de la Bella Durmiente diseñada se une específicamente a las repeticiones invertidas del transposón.
La Bella Durmiente media en la transposición precisa de corte y pegaje en diversos tipos de células de vertebrados.
Se ha demostrado que la Bella Durmiente es un sistema activo de ADN-transposón.

Conclusiones:

La Bella Durmiente es un sistema funcional de ADN-transposones de vertebrados.
Este sistema puede ser utilizado para la transformación genética y la mutagénesis por inserción.
La reconstrucción de antiguos transposones ofrece nuevas herramientas para la investigación genética.

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