The mammalian cerebral neocortex occupies the largest area of the cerebral cortex and is cytoarchitectually composed of six layers (I–VI). Recent molecular analysis has begun to reveal the existence of various developmental programs, including the genetic regulation of arealization of the neocortex. Although an increasing number of molecular determinants of the developmental stages of the neocortex have been identified, no genes specifically expressed in the adult neocortex have been identified to date. By global screening using microarrays, combined with systematic in situ hybridization, we identified a zinc-finger type transcription factor, Fez1, which is expressed predominantly in the mouse adult neocortex. No other genes in the neocortex have been shown to date to have their expression with such high specificity. Using two-color in situ hybridization, we show that Fez1 is mainly expressed in cortical layers V and VI, not in γ-aminobutyric acid neurons but in pyramidal neurons, the projection neurons of the cerebral cortex. Immunohistochemistry also shows that Fez1 is expressed in deep layers of the neocortex. Fez1 will be invaluable not only for the molecular understanding of corticogenesis but also for understanding the physiological functions of the adult neocortex, as well as for the use of its promoter in gene-manipulated animals and in conditional expression systems.