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 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.
Takada
Shuji
S
Division of Functional Genomics, Jichi Medical University, 3311-1 Yakushiji, Shimotsukeshi, Tochigi 329-0498, Japan.
Berezikov
Eugene
E
Yamashita
Yoshihiro
Y
Lagos-Quintana
Mariana
M
Kloosterman
Wigard P
WP
Enomoto
Munehiro
M
Hatanaka
Hisashi
H
Fujiwara
Shin-ichiro
S
Watanabe
Hideki
H
Soda
Manabu
M
Choi
Young Lim
YL
Plasterk
Ronald H A
RH
Cuppen
Edwin
E
Mano
Hiroyuki
H
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004622:Embryo, Mammalian / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D035683:MicroRNAs / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D014018:Tissue Distribution
D035683:MicroRNAs