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In contrast, functional annotations are available only for a very small fraction of these miRNAs, and even in these cases only partially.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>We developed a general Bayesian method for the inference of miRNA target sites, in which, for each miRNA, we explicitly model the evolution of orthologous target sites in a set of related species. Using this method we predict target sites for all known miRNAs in flies, worms, fish, and mammals. By comparing our predictions in fly with a reference set of experimentally tested miRNA-mRNA interactions we show that our general method performs at least as well as the most accurate methods available to date, including ones specifically tailored for target prediction in fly. An important novel feature of our model is that it explicitly infers the phylogenetic distribution of functional target sites, independently for each miRNA. This allows us to infer species-specific and clade-specific miRNA targeting. We also show that, in long human 3' UTRs, miRNA target sites occur preferentially near the start and near the end of the 3' UTR.<\/jats:p>\n            <jats:p>To characterize miRNA function beyond the predicted lists of targets we further present a method to infer significant associations between the sets of targets predicted for individual miRNAs and specific biochemical pathways, in particular those of the KEGG pathway database. We show that this approach retrieves several known functional miRNA-mRNA associations, and predicts novel functions for known miRNAs in cell growth and in development.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>We have presented a Bayesian target prediction algorithm without any tunable parameters, that can be applied to sequences from any clade of species. The algorithm automatically infers the phylogenetic distribution of functional sites for each miRNA, and assigns a posterior probability to each putative target site. The results presented here indicate that our general method achieves very good performance in predicting miRNA target sites, providing at the same time insights into the evolution of target sites for individual miRNAs. Moreover, by combining our predictions with pathway analysis, we propose functions of specific miRNAs in nervous system development, inter-cellular communication and cell growth. The complete target site predictions as well as the miRNA\/pathway associations are accessible on the ElMMo web server.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1471-2105-8-69","type":"journal-article","created":{"date-parts":[[2007,3,2]],"date-time":"2007-03-02T08:25:40Z","timestamp":1172823940000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":260,"title":["Inference of miRNA targets using evolutionary conservation and pathway analysis"],"prefix":"10.1186","volume":"8","author":[{"given":"Dimos","family":"Gaidatzis","sequence":"first","affiliation":[]},{"given":"Erik","family":"van Nimwegen","sequence":"additional","affiliation":[]},{"given":"Jean","family":"Hausser","sequence":"additional","affiliation":[]},{"given":"Mihaela","family":"Zavolan","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2007,3,1]]},"reference":[{"key":"1441_CR1","doi-asserted-by":"publisher","first-page":"843","DOI":"10.1016\/0092-8674(93)90529-Y","volume":"75","author":"R Lee","year":"1993","unstructured":"Lee R, Feinbaum R, Ambros V: The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. 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