Cloning and expression profiling of testis-expressed microRNAs

Seungil Ro, Chanjae Park, Kenton M. Sanders, John R. McCarrey, Wei Yan

Research output: Contribution to journalArticlepeer-review

202 Scopus citations

Abstract

Using a new small RNA cloning method, we identified 141 miRNAs from the mouse testis, of which 29 were novel. The 141 miRNAs were mapped onto all chromosomes except the Y chromosome and 2/3 of these miRNA genes exist as clusters. ∼ 70% of these miRNA genes were located in intronic or intergenic regions, whereas the remaining miRNAs were derived from exonic sequences. We further validated these cloned miRNAs by examining their expression in multiple mouse organs including developing testes and also in purified spermatogenic cells using semi-quantitative PCR analyses. Our expression profiling assays revealed that 60% of the testis-expressed miRNAs were ubiquitously expressed and the remaining are either preferentially (35%) or exclusively (5%) expressed in the testis. We also observed a lack of strand selection during testicular miRNA biogenesis, characterized by paired expression of both the 5′ strands and 3′ strands derived from the same precursor miRNAs. The present work identified numerous miRNAs preferentially or exclusively expressed in the testis, which would be interesting targets for further functional studies.

Original languageEnglish (US)
Pages (from-to)592-602
Number of pages11
JournalDevelopmental Biology
Volume311
Issue number2
DOIs
StatePublished - Nov 15 2007
Externally publishedYes

Keywords

  • Germ cells
  • microRNAs
  • Non-coding RNAs
  • Small RNAs
  • Spermatogenesis
  • Testis

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

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