MicroRNAs play pivotal roles in gene regulation. Despite various research efforts on microRNAs, how microRNA target genes are transcriptionally regulated and how the transcriptional regulation of microRNA target genes relates to that of the microRNA genes are not well studied. By investigating the transcriptional regulation of microRNA target genes, we found that different groups of target genes of the same microRNA are co-expressed under different conditions, and these groups rarely overlap with each other for the majority of microRNAs. We also discovered that co-expressed microRNA target genes are often co-regulated, and different groups of target genes of the same microRNA are often regulated differently. In addition, we observed that transcription factors regulating a microRNA gene often regulate its target genes. Our study sheds light on the regulation of microRNA target genes, which will facilitate the prediction of microRNA target genes and the understanding of the transcriptional regulation of microRNA genes. © 2011 Elsevier Inc.