comparative sequence analysis reveals an intricate network among rest creb and mirna in mediating neuronal gene expression background two distinct classes of regulators have been implicated in regulating neuronal gene expression and mediating neuronal identity transcription factors such as restnrsf re1 silencing transcription factor and creb camp response elementbinding protein and micrornas mirnas how these two classes of regulators act together to mediate neuronal gene expression is unclearresults using comparative sequence analysis here we report the identification of 895 sites nrse as the putative targets of rest a set of the identified nrse sites is present in the vicinity of the mirna genes that are specifically expressed in brainrelated tissues suggesting the transcriptional regulation of these mirnas by rest we have further identified target genes of these mirnas and discovered that rest and its cofactor complex are targets of multiple brainrelated mirnas including mir124a mir9 and mir132 given the role of both rest and mirna as repressors these findings point to a doublenegative feedback loop between rest and the mirnas in stabilizing and maintaining neuronal gene expression additionally we find that the brainrelated mirna genes are highly enriched with evolutionarily conserved camp response elements cre in their regulatory regions implicating the role of creb in the positive regulation of these mirnasconclusion the expression of neuronal genes and neuronal identity are controlled by multiple factors including transcriptional regulation through rest and posttranscriptional modification by several brainrelated mirnas we demonstrate that these different levels of regulation are coordinated through extensive feedbacks and propose a network among rest creb proteins and the brainrelated mirnas as a robust program for mediating neuronal gene expression