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P015 Extensive post-transcriptional regulation of miRNAs within embryonic stem cells Rupa Priti Sarkar, Geok Tan and Nick Dibb Imperial College London, London, United Kingdom MiRNAs are small non-coding RNAs that regulate diverse biological functions including cell proliferation, apoptosis and differentiation. Consequently it is not surprising that disruption of miRNA synthesis plays an important role in various human diseases including cancer. The biogenesis of miRNAs lends itself to regulation at several stages, from transcription of the miRNA gene to post-transcriptional control of primary, precursor and mature miRNA transcripts. We have analysed miRNAs that are uniquely expressed in either human embryonic stem cells (ESCs) or differentiated neuronal progenitors (NPCs). We have found that within ESCs miRNAs can be regulated transcriptionally and post-transcriptionally opposed to NPCs where miRNAs are regulated by transcription only. MiRNAs with high expression in ESCs and NPCs were investigated and out of these <40% of miRNA genes in ESCs are transcribed but not processed to maturation; whereas in NPCs all transcribed miRNAs genes are processed into mature miRNAs. Flexible regulation of miRNAs in ESCs and strict transcriptional regulation of miRNAs within NPCs may play a role in maintaining pluripotency and aiding differentiation. MiRNAs can be found in clusters sharing a single promoter, despite this, individual miRNAs within clusters are often differentially expressed. We have evidence of extensive alternative 3’polyadenylation of primary miRNA transcripts that isolate individual miRNA within clusters. Potentially alternative 3’polyadenylation may help regulate expression of a single miRNA within a cluster, allowing fine tuning of miRNA expression.