We cloned the 5'-flanking region of the mouse homolog of the Delta gene (Dill) and demonstrated that the sequence between nucleotide position -514 and -484 in the 5'-flanking region of D111 played a critical role in the regulation of its tissue-specific expression in neural stem cells (NSCs). Further, we showed that multiple POU-binding motifs, located within this short sequence of 30 bp, were essential for transcriptional activation of D111 and also that multiple tissue-specific nuclear factors recognized these POU-binding motifs in various combinations through differentiation of NSCs. Thus, POU-binding factors may play an important role in D111 expression in developing NSCs. (C) 2004 Elsevier Inc. All rights reserved.