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.