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Control basado en redox de la transformación y activación de los complejos de siRNA en entornos extracelulares

Burcu S Aytar¹, John P E Muller, Yukishige Kondo

¹Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.

Journal of the American Chemical Society

|May 25, 2013

Resumen

Los lípidos catiónicos redox-activos, el bis(11-ferrocenilundecilo) bromuro de dimetilamonio (BFDMA), permiten el control de la actividad lipopléxica del ARN pequeño interferente (siRNA). El BFDMA reducido promueve el silenciamiento de genes, mientras que el BFDMA oxidado puede activarse para la entrega controlada de siRNA.

Área de la Ciencia:

Química biofísica es la química biofísica.
Nanotecnología La nanotecnología es la nanotecnología.
Biología molecular La biología molecular.

Sus antecedentes:

Los lípidos catiónicos son cruciales para formar lipopléjicos con ácidos nucleicos como el siRNA para el silenciamiento de genes.
Los grupos de ferrocenilo redox-activos en el bis(11-ferrocenilundecilo) bromuro de dimetilamonio (BFDMA) ofrecen potencial para propiedades de lipoplejo sintonizables.

Objetivo del estudio:

Investigar las características físicas y la actividad biológica de los lipopléxidos de siRNA formados con el lípido catiónico redox-activo BFDMA.
Para determinar si el estado redox de BFDMA influye en las propiedades de lipoplex y la eficiencia de entrega de siRNA.

Principales métodos:

Complejación de siRNA con BFDMA para formar lipopléjicos.
Caracterización de las propiedades físicas del lipoplex, incluida la nanoestructura (cryo-TEM) y el potencial ζ.
Evaluación de la actividad de silenciamiento de genes en entornos celulares.
Activación in situ de lipopléxidos oxidados utilizando agentes reductores químicos.

Principales resultados:

Lipoplexes de siRNA con BFDMA reducido demostraron alto silenciamiento de genes específicos de la secuencia.
Los lipopléjicos con BFDMA oxidado no mostraron silenciamiento genético, lo que se correlaciona con una internalización celular deficiente.
Los lipopléxidos oxidados de BFDMA podrían ser activados por un agente reductor para inducir el silenciamiento de genes.
Se observaron diferencias de potencial de nanoestructura y ζ dependientes de redox.
Los mecanismos de control de redox observados con el ADN del plásmido se mantuvieron con siRNA.

Conclusiones:

El estado redox de BFDMA proporciona un mecanismo para controlar la actividad y la entrega de lipopléxidos cargados de siRNA.
La activación externa de los lipopléxidos ofrece una estrategia para la entrega controlada de siRNA en los sistemas celulares.
La capacidad de respuesta redox de BFDMA presenta nuevas oportunidades para el diseño de sistemas avanzados de entrega de siRNA.

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