In Vivo Inhibition of microRNA (miR) Using a Novel miR Inhibitor System Reveals a Role for miR‐17‐92 in Palate Elevation and/or Extension

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MicroRNAs (miRs) are short non‐coding RNA molecules approximately 22 nucleotides long. miRs bind to complementary targets on the 3′ untranslated region (UTR) of messenger RNAs (mRNAs), attenuating mRNA translation via either mRNA strand degradation or sequestration. Given their redundancy throughout the genome, conventional knockout strategies are difficult to employ when studying miRs. To circumvent this difficulty, we have developed a novel method of miR inhibition, the Plasmid‐Based miRNA Inhibition System (PMIS), to allow for the simultaneous knockdown of homologous miRs. Short hairpin structures that flank the anti‐sense miR target site on the PMIS inhibitor complex (PMIS‐IC) dramatically increase miR inhibition by associating with RISC complex factors. The PMIS‐IC is composed of native, unmodified nucleic acids and can be stably integrated into the genome. In this study, we employ this strategy to analyze the effects of inhibition of mature miRs in the miR‐17‐92 cluster. Here we show how the PMIS system can be used to inhibit miRs from the miR‐17‐92 cluster and associated family members in stable cell lines and transgenic mice. We also show how inhibition of the miR‐17‐92 cluster can lead to differences in the point of palate formation arrest in mice carrying PMIS‐ICs for miR‐17‐92. These results demonstrate the effectiveness of this novel miR inhibition strategy and indicate new roles for miR‐17‐92 in palatogenesis.Support or Funding InformationUniversity of Iowa Carver College of MedicineUniversity of Iowa College of DentistryUniversity of Iowa Research FoundationNational Institutes of Health (National Institute for Dental and Craniofacial Research)