Cited by 732

Mammalian microRNAs: experimental evaluation of novel and previously annotated genes

Chiang, H. Rosaria, Schoenfeld, Lori W., Ruby, J. Graham, Auyeung, Vincent C., Spies, Noah, Baek, Daehyun, Johnston, Wendy K., Russ, Carsten +6 more·2010·Genes & Development

Abstract

MicroRNAs (miRNAs) are small regulatory RNAs that derive from distinctive hairpin transcripts. To learn more about the miRNAs of mammals, we sequenced 60 million small RNAs from mouse brain, ovary, testes, embryonic stem cells, three embryonic stages, and whole newborns. Analysis of these sequences confirmed 398 annotated miRNA genes and identified 108 novel miRNA genes. More than 150 previously annotated miRNAs and hundreds of candidates failed to yield sequenced RNAs with miRNA-like features. Ectopically expressing these previously proposed miRNA hairpins also did not yield small RNAs, whereas ectopically expressing the confirmed and newly identified hairpins usually did yield small RNAs with the classical miRNA features, including dependence on the Drosha endonuclease for processing. These experiments, which suggest that previous estimates of conserved mammalian miRNAs were inflated, provide a substantially revised list of confidently identified murine miRNAs from which to infer the general features of mammalian miRNAs. Our analyses also revealed new aspects of miRNA biogenesis and modification, including tissue-specific strand preferences, sequential Dicer cleavage of a metazoan precursor miRNA (pre-miRNA), consequential 5′ heterogeneity, newly identified instances of miRNA editing, and evidence for widespread pre-miRNA uridylation reminiscent of miRNA regulation by Lin28.

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Research works printed but not linked: 5

  1. Blow MJ, Grocock RJ, van Dongen S, Enright AJ, Dicks E, Futreal PA, Wooster R, Stratton MR. 2006. RNA editing of human microRNAs. Genome Biol 7: R27. doi: 10.1186/gb-2006-7-4-r27. · DOI
  2. Griffiths-Jones S. 2004. The microRNA registry. Nucleic Acids Res 32: D109-D111. doi: 10.1093/nar/gkh023. · DOI
  3. Kuhn RM, Karolchik D, Zweig AS, Wang T, Smith KE, Rose- nbloom KR, Rhead B, Raney BJ, Pohl A, Pheasant M, et al. 2009. The UCSC Genome Browser Database: Update 2009. Nucleic Acids Res 37: D755-D761. doi: 10.1093/nar/gkn875. · DOI
  4. Pruitt KD, Tatusova T, Maglott DR. 2005. NCBI Reference Sequence (RefSeq): A curated non-redundant sequence data- base of genomes, transcripts and proteins. Nucleic Acids Res 33: D501-D504. doi: 10.1093/nar/gki025. · DOI
  5. Wu H, Ye C, Ramirez D, Manjunath N. 2009. Alternative processing of primary microRNA transcripts by Drosha generates 59 end variation of mature microRNA. PLoS One 4: e7566. doi: 10.1371/journal.pone.0007566. · DOI

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Authors
Chiang, H. R., Schoenfeld, L. W., Ruby, J. G., Auyeung, V. C., Spies, N., Baek, D., Johnston, W. K., Russ, C., Luo, S., Babiarz, J. E., Blelloch, R., Schroth, G. P., Nusbaum, C., & Bartel, D. P.
Venue
Genes & Development
Published
2010-04-22
Volume
24
Issue
10
Pages
992-1009
ISSN
  • 0890-9369
  • 1549-5477
Cited by
732
References
7
Language
en

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