Cloning and expression profiling of testis-expressed microRNAs.
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. approximately 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.
Ro
Seungil
S
Department of Physiology and Cell Biology, University of Nevada School of Medicine, Anderson Biomedical Science Building 105C/111, 1664 North Virginia Street, MS 352, Reno, NV 89557, USA.
Park
Chanjae
C
Sanders
Kenton M
KM
McCarrey
John R
JR
Yan
Wei
W
D000818:Animals; D003001:Cloning, Molecular; D019295:Computational Biology; D020869:Gene Expression Profiling; D006801:Humans; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D015203:Reproducibility of Results; D013091:Spermatogenesis / Q000502:physiology*; D013737:Testis / Q000502:physiology*
D035683:MicroRNAs