Targeted in vivo genetic manipulation of the mouse or rat brain by in utero electroporation with a triple-electrode probe View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2016-02-04

AUTHORS

Joanna Szczurkowska, Andrzej W Cwetsch, Marco dal Maschio, Diego Ghezzi, Gian Michele Ratto, Laura Cancedda

ABSTRACT

This protocol is an extension to:Nat. Protoc. 1, 1552-1558 (2006); doi:10.1038/nprot.2006.276; published online 9 November 2006This article describes how to reliably electroporate with DNA plasmids rodent neuronal progenitors of the hippocampus; the motor, prefrontal and visual cortices; and the cerebellum in utero. As a Protocol Extension article, this article describes an adaptation of an existing Protocol and offers additional applications. The earlier protocol describes how to electroporate mouse embryos using two standard forceps-type electrodes. In the present protocol, additional electroporation configurations are possible because of the addition of a third electrode alongside the two standard forceps-type electrodes. By adjusting the position and polarity of the three electrodes, the electric field can be directed with great accuracy to different neurogenic areas. Bilateral transfection of brain hemispheres can be achieved after a single electroporation episode. Approximately 75% of electroporated embryos survive to postnatal ages, and depending on the target area, 50-90% express the electroporated vector. The electroporation procedure takes 1 h 35 min. The protocol is suitable for the preparation of animals for various applications, including histochemistry, behavioral studies, electrophysiology and in vivo imaging. More... »

PAGES

399-412

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nprot.2016.014

DOI

http://dx.doi.org/10.1038/nprot.2016.014

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1040500166

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/26844428


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