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MicroRNAs in search of a target.

Cold Spring Harbor symposia on quantitative biology

metabolism, genetics, RNA-Induced Silencing Complex, Nuclear Proteins, MicroRNAs, Humans, Genes, Helminth, Gene Expression Regulation, Computational Biology, Caenorhabditis elegans Proteins, Caenorhabditis elegans, Animals

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      Abstract

      As the number of known microRNAs (miRNAs) increases, and their importance in physiology and disease becomes apparent, the identification of their regulatory targets is a requisite for a full characterization of their biological functions. Computational methods based on sequence homology and phylogenetic conservation have spearheaded this effort in the last 3 years, but they may not be sufficient. Experimental studies are now needed to extend and validate the computational predictions and further our understanding of target recognition by miRNAs.

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      Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets.

      We predict regulatory targets of vertebrate microRNAs (miRNAs) by identifying mRNAs with conserved complementarity to the seed (nucleotides 2-7) of the miRNA. An overrepresentation of conserved adenosines flanking the seed complementary sites in mRNAs indicates that primary sequence determinants can supplement base pairing to specify miRNA target recognition. In a four-genome analysis of 3' UTRs, approximately 13,000 regulatory relationships were detected above the estimate of false-positive predictions, thereby implicating as miRNA targets more than 5300 human genes, which represented 30% of our gene set. Targeting was also detected in open reading frames. In sum, well over one third of human genes appear to be conserved miRNA targets.
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        Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs.

        MicroRNAs (miRNAs) are a class of noncoding RNAs that post-transcriptionally regulate gene expression in plants and animals. To investigate the influence of miRNAs on transcript levels, we transfected miRNAs into human cells and used microarrays to examine changes in the messenger RNA profile. Here we show that delivering miR-124 causes the expression profile to shift towards that of brain, the organ in which miR-124 is preferentially expressed, whereas delivering miR-1 shifts the profile towards that of muscle, where miR-1 is preferentially expressed. In each case, about 100 messages were downregulated after 12 h. The 3' untranslated regions of these messages had a significant propensity to pair to the 5' region of the miRNA, as expected if many of these messages are the direct targets of the miRNAs. Our results suggest that metazoan miRNAs can reduce the levels of many of their target transcripts, not just the amount of protein deriving from these transcripts. Moreover, miR-1 and miR-124, and presumably other tissue-specific miRNAs, seem to downregulate a far greater number of targets than previously appreciated, thereby helping to define tissue-specific gene expression in humans.
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          The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14

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            Author and article information

            Journal
            10.1101/sqb.2006.71.032
            17381288

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