Direct and efficient transfection of mouse neural stem cells and mature neurons by in vivo mRNA electroporation [TECHNIQUES AND RESOURCES ARTICLE]

Stephane Bugeon, Antoine de Chevigny, Camille Boutin, Natalie Core, Stefan Wild, Andreas Bosio, Harold Cremer, and Christophe Beclin

In vivo brain electroporation of DNA expression vectors is a widely used method for lineage and gene function studies in the developing and postnatal brain. However, transfection efficiency of DNA is limited and adult brain tissue is refractory to electroporation. Here we present a systematic study of mRNA as a vector for acute genetic manipulation in the developing and adult brain. We demonstrate that mRNA electroporation is far more efficient than DNA and leads to faster and more homogeneous protein expression in vivo. Importantly, mRNA electroporation allows the manipulation of neural stem cells and postmitotic neurons in the adult brain with minimal invasive procedures. Finally, we show that this approach can be efficiently used for functional studies as exemplified by transient overexpression of the neurogenic factor Myt1l and by stably inactivating Dicer nuclease in vivo in adult born olfactory bulb interneurons and in fully integrated cortical projection neurons.

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