1/10 HH 5 6-1 I II. NEDO 2/10 sirna sirna RNA mirna RNA non-coding RNA 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 RNA Noncoding RNARNARNA)
3/10 4/10
5/10 RNA 6/10
7/10 RNA BI RNA RNA RNA RNA ncrna ncrna mirna 8/10
PL 1: 400 350 300 250 200 150 100 50 0 9/10 2005(H17) 2006(H18) 2007(H19) 2008(H20) 2009(H21) 10/10 RNA RNA RNA RNA ips NEDO
1/42 HH 5 6- III. IV. 2/42
3/42 4/53 4/42
5/42 dataset2: length 50-500 dataset3: length 500-1500 +10%, +10% +30%, +30% Hamada M et al., Bioinformatics 25(12), 2009 (ISMB 2009) 6/42 Stemloc) 1007.2 MXSCARNA 1001 500RNA 1 Kiryu H et al., Bioinformaitcs 23(13), 2007 17 Tabei Y et al., Bioinformatics 22(14), 2006 Tabei Y et al., BMC Bioinformatics 9:33, 2008
7/42 8/42
9/42 mirnamirbase mirbase A B A A A B B B 2,103 5,538 155 Human Accelerated Region Ultra Conserved Region 1433 Ultra Stable Fold Inverted Duplication RNA 10/42 AU GA AG GC UA 680 RNA mirrim 510 RNA1500
11/42 12/42 RNA 107,707 RNA 2,325,800 182,638 Kin T et al., Nucleic Acids Res. 35 (DB), 2007 Kin T et al., Bioinformatics 23(21), 2007 Mituyama T et al., Nucleic Acids Res. 37 (DB), 2009
13/42 14/42
15/42 16/42
17/42 18/42 MS FTMS
RNA 19/42 ) tensity (cps) Int Time (min) (30 ) x mhen1 Relativeabundance 100 mgld-2 (+/-) 90 22 nt 80 70 21 nt 23 nt (3'-U) 60 50 40 30 23 nt (3'-A) Relativeabundance 100 mgld-2 (-/-) 90 21 nt 22 nt 80 70 60 23 nt (3'-U) 50 40 30 HO O N O NH2 N HO OCH 3 2 -O-methylcytidine pirna mhen1 in situ hybridization 20 10 0 30.5 31.0 31.5 32.0 32.5 Time (min) 21mer 5' 5' 3' 22mer Deguanylation 3' 5' UUU Degradation 20 23 nt (3'-A) 10 0 32.0 33.0 34.0 35.0 Time (min) UUUG 3' Shortening & Degradation Uridylation Deuridylation Adenylation n by GLD D2 Deaden enylation 23mer (3'-U) 5' UUUGU 3' 23mer (3'-A) 3' 5' UUUGA Elongation & Stabilization 20/42
21/42 22/42
23/42 24/42
mirna 25/42 1998 2001 2004 2009 RNAi mirna RNA 2005 ncrna PJ mirna ncrna a ncrna RNA / RNA ncrna ncrna mirna 26/42 >1300miRNA (2006.12) mirna
27/42 Cell viability activity Caspase 3/7 a 1.2 1 0.8 0.6 0.4 0.2 0 5.0 4.0 3.0 2.0 1.0 0.0 Cell viability Caspase 3/7 activity le et-7a mir- R-15a mir R-16 mir- R-143 mir- R-145 mir-a ir-b mi ir-c mi ir-d mi mir-e mir-nega acon [mirna] =25nM Tumor size (mm3) 700 600 500 400 300 200 100 0 control mir-d mir-c 0 7 10 13 16 Days after injection ips mirna 28/42 A Num mberofh heslike colonie es B C OSK+hmir302siPS hes IIItubulin Sox17 SMA
29/42 ncrna 2% 93% noncoding RNA Noncoding RNA = RNA = dark matter ncrna 30/42 II > I >70ncRNA ncrnanmd (Ideueetal.,Genes&Dev2007) 60ncRNA (Hokiietal.,Gene2006) 100ncRNA (Sasakietal.,BBRC2007a,b) 10 RNA (Kikuchietal.,NAR2009)
ncrnarna RNAi ASO 31/42 RNaseH RNA mrna Antisense chimera mrna-like ncrna ASO sirna Small nuclear ncrna 6 24 6 24 6 24 6 24 U84 50ncRNA10 2009-171895 Ideue et al., RNA 2009 ncrna 32/42 MEN / ncrna MEN / : architectural RNA PSF PSP1 MEN MEN p54 Sasaki et al., PNAS 2009
33/42 34/42
RNAi 35/42 mirna sirna esirna (Nature 2008) AGO2 pirna (G&D 2006, Science 2007 Nature 2009) mirna or sirna mirna or sirna mirna or sirna mirna or sirna Ago AgomiRNA RISC ncrna 36/42
37/42 38/42
39/42 40/42
41/42 42/42 mirna mirna mirnaips ncrnarna