Mouse microRNA profiles determined with a new and sensitive cloning method

MicroRNAs (miRNAs) are noncoding RNA molecules of 21 to 24 nt that regulate the expression of target genes in a post-transcriptional manner. Although evidence indicates that miRNAs play essential roles in embryogenesis, cell differentiation and pathogenesis of human diseases, extensive miRNA profili...

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Bibliographic Details
Published in:Nucleic acids research 2006-10, Vol.34 (17), p.e115-e115
Main Authors: Takada, Shuji, Berezikov, Eugene, Yamashita, Yoshihiro, Lagos-Quintana, Mariana, Kloosterman, Wigard P, Enomoto, Munehiro, Hatanaka, Hisashi, Fujiwara, Shin-ichiro, Watanabe, Hideki, Soda, Manabu, Choi, Young Lim, Plasterk, Ronald H A, Cuppen, Edwin, Mano, Hiroyuki
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Cloning, Molecular - methods
Embryo, Mammalian - metabolism
Gene Expression Profiling - methods
Humans
Jurkat Cells
Methods Online
Mice
MicroRNAs - genetics
MicroRNAs - isolation & purification
MicroRNAs - metabolism
Molecular Sequence Data
Polymerase Chain Reaction - methods
Tissue Distribution
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Summary:MicroRNAs (miRNAs) are noncoding RNA molecules of 21 to 24 nt that regulate the expression of target genes in a post-transcriptional manner. Although evidence indicates that miRNAs play essential roles in embryogenesis, cell differentiation and pathogenesis of human diseases, extensive miRNA profiling in cells or tissues has been hampered by the lack of sensitive cloning methods. Here we describe a highly efficient profiling method, termed miRNA amplification profiling (mRAP), as well as its application both to mouse embryos at various developmental stages and to adult mouse organs. A total of 77,436 Small-RNA species was sequenced, with 11,776 of these sequences found to match previously described miRNAs. With the use of a newly developed computational prediction algorithm, we further identified 229 independent candidates for previously unknown miRNAs. The expression of some of these candidate miRNAs was confirmed by northern blot analysis and whole-mount in situ hybridization. Our data thus indicate that the total number of miRNAs in vertebrates is larger than previously appreciated and that the expression of these molecules is tightly controlled in a tissue- and developmental stage-specific manner.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/gkl653