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In ovo Electroporation of Chicken Embryos

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Electroporation is a technique used in biomedical research that allows for the manipulation of gene expression via the delivery of foreign genetic material into cells. More specifically, in ovo electroporation is performed on early developing chicks (Gallus gallus domesticus) contained within their eggshells. In this procedure, DNA or knockdown constructs are first injected into a target tissue. However, the genetic material is unable to penetrate the plasma membrane to carry out its function within the cell. To solve this problem, an electrical field is applied, causing temporary disruptions to membrane stability. This electric field also causes the negatively charged nucleic acids to migrate toward the positively charged electrode through the holes in the plasma membrane, thus effectively driving the DNA or knockdown construct into the cell. The major advantage of this technique is that the delivery of genetic material can be localized to isolated cell types at specific developmental time points. As a result, the genetic mechanisms that govern individual developmental events can be examined.

This video provides an overview of the principles behind in ovo electroporation and introduces the tools required for the technique, including capillary needles, electrodes, and an electroporator. A step-by-step protocol for carrying out the procedure is also presented prior to discussion of a few fascinating examples of how the technique is used to perform a variety of genetic manipulations in chicken embryos.

Procedura

Electroporation is a technique used to introduce foreign genetic material into cells. Electroporation of chick embryos inside the egg, or in ovo electroporation, is a valuable tool for developmental biologists because the delivery of genetic material can be localized to specific tissues and specific developmental time points. This video will introduce the basic principles of in ovo electroporation, describe essential steps of the procedure, and discuss how this technique can be used to study biological p

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Tags

ElectroporationIn Ovo ElectroporationGenetic MaterialCellsChick EmbryosDevelopmental BiologistsTissuesDevelopmental Time PointsDNA DeliveryMicroinjectionPlasma MembraneElectrical CurrentPoresPositive ElectrodeTransfected CellsHamburger Hamilton Stage 20Ex Ovo Culture

Vai a...

0:00

Overview

0:46

Basic Principles of Electroporation

1:37

Preparing for Electroporation

3:58

Performing the Electroporation

4:45

Applications

6:21

Summary

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