In utero and ex vivo Electroporation for Gene Expression in Mouse Retinal Ganglion Cells

Timothy J Petros1, Alexandra Rebsam1, Carol A Mason2, 1
1Departments of Pathology and Cell Biology, and Neuroscience, Columbia University College of Physicians and Surgeons, 2Department of Ophthalmology, Columbia University College of Physicians and Surgeons

Here we present two techniques for manipulating gene expression in murine retinal ganglion cells (RGCs) by in utero and ex vivo electroporation. These techniques enable one to examine how alterations in gene expression affect RGC development, axon guidance, and functional properties.

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In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos

William Walantus, Laura Elias, Arnold Kriegstein
Institute for Regeneration Medicine, University of California, San Francisco

In-utero in-vivo injection and electroporation of the embryonic rat neocortex provides a powerful tool for the manipulation of individual progenitors lining the walls of the lateral ventricle. This technique is now widely used to study the processes involved in corticogenesis by over-expressing or knocking down genes and observing the effects on cellular proliferation, migration, and differentiation. In comparison to traditional knockout strategies, in-utero electroporation provides a rapid means to manipulate a population of cells during a specific temporal window. In this video protocol, we outline the experimental methodology for preparing rats for surgery, exposing the uterine horns through laporatomy, injecting DNA into the lateral ventricles of the developing embryo, electroporating DNA into the progenitors lining the lateral wall, and caring for animals post-surgery. Our laboratory uses this protocol for surgeries on E15-E21 rats, however it is most commonly performed at E16 as shown in this video.

In Ovo Electroporations of HH Stage 10 Chicken Embryos

Marissa C. Blank1, Victor Chizhikov2, Kathleen J. Millen2
1Department of Molecular Genetics and Cell Biology, University of Chicago, 2Department of Human Genetics, University of Chicago

Chick in ovo electroporation is a technique which allows genetic manipulation of the avian embryo. Common applications of this technique include functional analysis of genes and putative enhancer elements. This video demonstrates neural tube electroporation in HH 10 chick embryos. Injection technique and proper egg handling are discussed.

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