The chicken embryo has served as a classic model system for developmental studies due to its easy access for surgical manipulations and a wealth of data about chicken embryogenesis. Notably, the mechanisms controlling limb development have been explored best in the chick. Recently, the method of in ovo electroporation has been used successfully to transfect particular cells/tissues during embryonic development, without the production or infectivity associated with retroviruses. With the sequencing of the chicken genome near completion, this approach will provide a powerful opportunity to examine the function of chicken genes and their counterparts in other species. In ovo electroporation has been most effectively used to date for ectopic or overexpression analyses. However, recent studies indicate that this approach can be used successfully for loss-of-function analyses, including protein knockdown experiments with morpholinos and RNAi. Here, I will discuss parameters for using in ovo electroporation successfully to study developmental processes. Developmental Dynamics 229:433–439, 2004. © 2004 Wiley-Liss, Inc.