Induced pluripotent stem cells (iPSCs) provide a powerful and novel approach to generating transgenic animals by enabling complex genetic manipulations. iPSCs from various species have been derived by introducing different combinations of reprogramming factors. However, chicken iPSCs have not been yet reported. In this study, we describe the generation of chicken induced pluripotent stem cells (ciPSCs) by transfection of a nonviral minicircle DNA construct into chicken embryonic fibroblast cells (CEFs). Following transfection, ciPSC colonies showed stem cell morphology. They expressed pluripotent markers such as POU5F1, SOX2, NANOG, SSEA-1. ciPSCs were capable of differentiate into 3 germ layers in vitro and in vivo. ciPSCs were capable of incorporating into tissues from all three germ layers of the chimeric chickens. This research demonstrates for the first time that ciPSCs may be a robust tool for the generation of transgenic animals with unique and specialized traits.