Oct4 is one of the most important transcription factors required to maintain an undifferentiated state (self-renewal) and pluripotency of human and mouse embryonic stem (ES) cells as well as early embryonic cells. In addition, Oct4 is the only known transcription factor that has to be exogenously introduced into differentiated cells to make induced pluripotent stem (iPS) cells. Therefore, it is of great importance to understand how Oct4 transcription is regulated in ES cells and embryos and how it becomes activated during iPS cell formation. In this article, we will review the regulation of the mouse Oct4 gene from the viewpoint of DNA methylation, binding of orphan nuclear receptors, histone modifications and synergistic effects with other pluripotency factors. We will also raise several key questions that need to be addressed in future work to improve our understanding of Oct4 gene regulation and its essential role in self-renewal and pluripotency.