Vertebrate development culminates in the generation of proper proportions of a large variety of different cell types and subtypes essential for tissue, organ and system functions in the right place at the right time. Foxn4, a member of the forkhead box/winged-helix transcription factor superfamily, is expressed in mitotic progenitors and/or postmitotic precursors in both neural (e.g., retina and spinal cord) and non-neural tissues (e.g., atrioventricular canal and proximal airway). During development of the central nervous system, Foxn4 is required to specify the amacrine and horizontal cell fates from multipotent retinal progenitors while suppressing the alternative photoreceptor cell fates through activating Dll4-Notch signaling. Moreover, it activates Dll4-Notch signaling to drive commitment of p2 progenitors to the V2b and V2c interneuron fates during spinal cord neurogenesis. In development of non-neural tissues, Foxn4 plays an essential role in the specification of the atrioventricular canal and is indirectly required for patterning the distal airway during lung development. In this review, we highlight current understanding of the structure, expression and developmental functions of Foxn4 with an emphasis on its cell-autonomous and non-cell-autonomous roles in different tissues and animal model systems.