JP2010528667A - トリの生産形質の改変 - Google Patents
トリの生産形質の改変 Download PDFInfo
- Publication number
- JP2010528667A JP2010528667A JP2010511446A JP2010511446A JP2010528667A JP 2010528667 A JP2010528667 A JP 2010528667A JP 2010511446 A JP2010511446 A JP 2010511446A JP 2010511446 A JP2010511446 A JP 2010511446A JP 2010528667 A JP2010528667 A JP 2010528667A
- Authority
- JP
- Japan
- Prior art keywords
- seq
- nucleic acid
- acid molecule
- gene
- shrna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 241000271566 Aves Species 0.000 title claims abstract description 27
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 26
- 238000012986 modification Methods 0.000 title description 6
- 230000004048 modification Effects 0.000 title description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims abstract description 47
- 238000000034 method Methods 0.000 claims abstract description 47
- 241000287828 Gallus gallus Species 0.000 claims abstract description 34
- 108020004459 Small interfering RNA Proteins 0.000 claims abstract description 34
- 102000040650 (ribonucleotides)n+m Human genes 0.000 claims abstract description 22
- 150000007523 nucleic acids Chemical class 0.000 claims description 100
- 102000039446 nucleic acids Human genes 0.000 claims description 96
- 108020004707 nucleic acids Proteins 0.000 claims description 96
- 108091027967 Small hairpin RNA Proteins 0.000 claims description 68
- 239000004055 small Interfering RNA Substances 0.000 claims description 55
- 108090000623 proteins and genes Proteins 0.000 claims description 49
- 239000013598 vector Substances 0.000 claims description 39
- 235000013330 chicken meat Nutrition 0.000 claims description 32
- 239000002773 nucleotide Substances 0.000 claims description 32
- 125000003729 nucleotide group Chemical group 0.000 claims description 28
- 210000003205 muscle Anatomy 0.000 claims description 19
- 102000004169 proteins and genes Human genes 0.000 claims description 19
- 108010056852 Myostatin Proteins 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 13
- 241000286209 Phasianidae Species 0.000 claims description 9
- 230000009467 reduction Effects 0.000 claims description 8
- 241000272517 Anseriformes Species 0.000 claims description 7
- 101150078435 dmrt1 gene Proteins 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 5
- 239000007924 injection Substances 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 3
- 210000004027 cell Anatomy 0.000 description 59
- 230000014509 gene expression Effects 0.000 description 49
- 239000013612 plasmid Substances 0.000 description 26
- 235000013601 eggs Nutrition 0.000 description 22
- 230000001629 suppression Effects 0.000 description 14
- 230000000692 anti-sense effect Effects 0.000 description 12
- 239000013604 expression vector Substances 0.000 description 12
- 101150048453 MSTN gene Proteins 0.000 description 11
- 230000004927 fusion Effects 0.000 description 11
- 239000013615 primer Substances 0.000 description 10
- 238000012228 RNA interference-mediated gene silencing Methods 0.000 description 9
- 230000009368 gene silencing by RNA Effects 0.000 description 9
- 238000013518 transcription Methods 0.000 description 9
- 230000035897 transcription Effects 0.000 description 9
- 238000004422 calculation algorithm Methods 0.000 description 8
- 238000003197 gene knockdown Methods 0.000 description 8
- 238000001890 transfection Methods 0.000 description 8
- 108020004414 DNA Proteins 0.000 description 7
- 102100039939 Growth/differentiation factor 8 Human genes 0.000 description 7
- 102000014450 RNA Polymerase III Human genes 0.000 description 7
- 108010078067 RNA Polymerase III Proteins 0.000 description 7
- 238000003556 assay Methods 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 241000588724 Escherichia coli Species 0.000 description 6
- 108091028043 Nucleic acid sequence Proteins 0.000 description 6
- 108020004999 messenger RNA Proteins 0.000 description 6
- 230000002441 reversible effect Effects 0.000 description 6
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 5
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 5
- 108700008625 Reporter Genes Proteins 0.000 description 5
- 238000013459 approach Methods 0.000 description 5
- 238000010276 construction Methods 0.000 description 5
- 210000002257 embryonic structure Anatomy 0.000 description 5
- 239000013613 expression plasmid Substances 0.000 description 5
- 239000012894 fetal calf serum Substances 0.000 description 5
- 238000000684 flow cytometry Methods 0.000 description 5
- 210000001161 mammalian embryo Anatomy 0.000 description 5
- 241000894007 species Species 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 241000283690 Bos taurus Species 0.000 description 4
- 102100030068 Doublesex- and mab-3-related transcription factor 1 Human genes 0.000 description 4
- 101100223985 Gallus gallus DMRT1 gene Proteins 0.000 description 4
- 101000864807 Homo sapiens Doublesex- and mab-3-related transcription factor 1 Proteins 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 108700026244 Open Reading Frames Proteins 0.000 description 4
- 210000004102 animal cell Anatomy 0.000 description 4
- 238000010367 cloning Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000002452 interceptive effect Effects 0.000 description 4
- 239000002502 liposome Substances 0.000 description 4
- 238000010369 molecular cloning Methods 0.000 description 4
- -1 pseudouracil Chemical compound 0.000 description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4
- 230000014616 translation Effects 0.000 description 4
- 102000002322 Egg Proteins Human genes 0.000 description 3
- 108010000912 Egg Proteins Proteins 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 101000886557 Gallus gallus Growth/differentiation factor 8 Proteins 0.000 description 3
- 108700011259 MicroRNAs Proteins 0.000 description 3
- 108091034117 Oligonucleotide Proteins 0.000 description 3
- 108091081021 Sense strand Proteins 0.000 description 3
- 210000004712 air sac Anatomy 0.000 description 3
- 125000002091 cationic group Chemical group 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000012761 co-transfection Methods 0.000 description 3
- 230000004069 differentiation Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 210000003746 feather Anatomy 0.000 description 3
- 210000002950 fibroblast Anatomy 0.000 description 3
- 238000003633 gene expression assay Methods 0.000 description 3
- 210000002149 gonad Anatomy 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 239000002679 microRNA Substances 0.000 description 3
- 230000001124 posttranscriptional effect Effects 0.000 description 3
- 244000144977 poultry Species 0.000 description 3
- 235000013594 poultry meat Nutrition 0.000 description 3
- 230000002103 transcriptional effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- LRFVTYWOQMYALW-UHFFFAOYSA-N 9H-xanthine Chemical compound O=C1NC(=O)NC2=C1NC=N2 LRFVTYWOQMYALW-UHFFFAOYSA-N 0.000 description 2
- 244000215068 Acacia senegal Species 0.000 description 2
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 229920002307 Dextran Polymers 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- 229920000084 Gum arabic Polymers 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 2
- 229930182816 L-glutamine Natural products 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 229930182555 Penicillin Natural products 0.000 description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical class OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 2
- 241000271567 Struthioniformes Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical class OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 101710137500 T7 RNA polymerase Proteins 0.000 description 2
- 108700029229 Transcriptional Regulatory Elements Proteins 0.000 description 2
- 108091026822 U6 spliceosomal RNA Proteins 0.000 description 2
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 2
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 2
- 235000010489 acacia gum Nutrition 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 210000004671 cell-free system Anatomy 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 239000013599 cloning vector Substances 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 230000001605 fetal effect Effects 0.000 description 2
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 230000002538 fungal effect Effects 0.000 description 2
- 101150004578 gdf-8 gene Proteins 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000005090 green fluorescent protein Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 229920000609 methyl cellulose Polymers 0.000 description 2
- 239000001923 methylcellulose Substances 0.000 description 2
- 235000010981 methylcellulose Nutrition 0.000 description 2
- 229940049954 penicillin Drugs 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 210000001236 prokaryotic cell Anatomy 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 239000012096 transfection reagent Substances 0.000 description 2
- 241001430294 unidentified retrovirus Species 0.000 description 2
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 1
- 101710175516 14 kDa zinc-binding protein Proteins 0.000 description 1
- KSXTUUUQYQYKCR-LQDDAWAPSA-M 2,3-bis[[(z)-octadec-9-enoyl]oxy]propyl-trimethylazanium;chloride Chemical compound [Cl-].CCCCCCCC\C=C/CCCCCCCC(=O)OCC(C[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC KSXTUUUQYQYKCR-LQDDAWAPSA-M 0.000 description 1
- WYDKPTZGVLTYPG-UHFFFAOYSA-N 2,8-diamino-3,7-dihydropurin-6-one Chemical compound N1C(N)=NC(=O)C2=C1N=C(N)N2 WYDKPTZGVLTYPG-UHFFFAOYSA-N 0.000 description 1
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 1
- UZOVYGYOLBIAJR-UHFFFAOYSA-N 4-isocyanato-4'-methyldiphenylmethane Chemical compound C1=CC(C)=CC=C1CC1=CC=C(N=C=O)C=C1 UZOVYGYOLBIAJR-UHFFFAOYSA-N 0.000 description 1
- OVONXEQGWXGFJD-UHFFFAOYSA-N 4-sulfanylidene-1h-pyrimidin-2-one Chemical compound SC=1C=CNC(=O)N=1 OVONXEQGWXGFJD-UHFFFAOYSA-N 0.000 description 1
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- DIHQLCHDZVJFCV-UHFFFAOYSA-N 5-(fluoromethyl)-1h-pyrimidine-2,4-dione Chemical compound FCC1=CNC(=O)NC1=O DIHQLCHDZVJFCV-UHFFFAOYSA-N 0.000 description 1
- SVXNJCYYMRMXNM-UHFFFAOYSA-N 5-amino-2h-1,2,4-triazin-3-one Chemical compound NC=1C=NNC(=O)N=1 SVXNJCYYMRMXNM-UHFFFAOYSA-N 0.000 description 1
- XZWMZFQOHTWGQE-UHFFFAOYSA-N 6-azathymine Chemical compound CC1=NNC(=O)NC1=O XZWMZFQOHTWGQE-UHFFFAOYSA-N 0.000 description 1
- CLGFIVUFZRGQRP-UHFFFAOYSA-N 7,8-dihydro-8-oxoguanine Chemical compound O=C1NC(N)=NC2=C1NC(=O)N2 CLGFIVUFZRGQRP-UHFFFAOYSA-N 0.000 description 1
- 108091034151 7SK RNA Proteins 0.000 description 1
- PFUVOLUPRFCPMN-UHFFFAOYSA-N 7h-purine-6,8-diamine Chemical compound C1=NC(N)=C2NC(N)=NC2=N1 PFUVOLUPRFCPMN-UHFFFAOYSA-N 0.000 description 1
- RGKBRPAAQSHTED-UHFFFAOYSA-N 8-oxoadenine Chemical compound NC1=NC=NC2=C1NC(=O)N2 RGKBRPAAQSHTED-UHFFFAOYSA-N 0.000 description 1
- MSSXOMSJDRHRMC-UHFFFAOYSA-N 9H-purine-2,6-diamine Chemical compound NC1=NC(N)=C2NC=NC2=N1 MSSXOMSJDRHRMC-UHFFFAOYSA-N 0.000 description 1
- 235000006491 Acacia senegal Nutrition 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000271560 Casuariidae Species 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- 241001137251 Corvidae Species 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- 101150074155 DHFR gene Proteins 0.000 description 1
- 108700029720 DMRT1 Proteins 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 241000271571 Dromaius novaehollandiae Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000160765 Erebia ligea Species 0.000 description 1
- 102000001690 Factor VIII Human genes 0.000 description 1
- 108010054218 Factor VIII Proteins 0.000 description 1
- 241000272184 Falconiformes Species 0.000 description 1
- 241000287227 Fringillidae Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 229930010555 Inosine Natural products 0.000 description 1
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000235648 Pichia Species 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 239000004372 Polyvinyl alcohol Substances 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 102100021557 Protein kinase C iota type Human genes 0.000 description 1
- 241000287530 Psittaciformes Species 0.000 description 1
- 102000009572 RNA Polymerase II Human genes 0.000 description 1
- 108010009460 RNA Polymerase II Proteins 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 241000220010 Rhode Species 0.000 description 1
- 108091028664 Ribonucleotide Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 108020004688 Small Nuclear RNA Proteins 0.000 description 1
- 102000039471 Small Nuclear RNA Human genes 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 108020004566 Transfer RNA Proteins 0.000 description 1
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 1
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 241000269370 Xenopus <genus> Species 0.000 description 1
- 101100277928 Xenopus laevis dmrt1-a gene Proteins 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 239000003429 antifungal agent Substances 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 210000000436 anus Anatomy 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229920001400 block copolymer Polymers 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000013553 cell monolayer Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000000412 dendrimer Substances 0.000 description 1
- 229920000736 dendritic polymer Polymers 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- FPAFDBFIGPHWGO-UHFFFAOYSA-N dioxosilane;oxomagnesium;hydrate Chemical compound O.[Mg]=O.[Mg]=O.[Mg]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O FPAFDBFIGPHWGO-UHFFFAOYSA-N 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical group P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 230000002222 downregulating effect Effects 0.000 description 1
- 210000003278 egg shell Anatomy 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000002969 egg yolk Anatomy 0.000 description 1
- 235000013345 egg yolk Nutrition 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 230000006718 epigenetic regulation Effects 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 150000002190 fatty acyls Chemical group 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 230000003325 follicular Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 235000003869 genetically modified organism Nutrition 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical class O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229960003786 inosine Drugs 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 101150066555 lacZ gene Proteins 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000001638 lipofection Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000000395 magnesium oxide Substances 0.000 description 1
- CPLXHLVBOLITMK-UHFFFAOYSA-N magnesium oxide Inorganic materials [Mg]=O CPLXHLVBOLITMK-UHFFFAOYSA-N 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- AXZKOIWUVFPNLO-UHFFFAOYSA-N magnesium;oxygen(2-) Chemical compound [O-2].[Mg+2] AXZKOIWUVFPNLO-UHFFFAOYSA-N 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 210000005075 mammary gland Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 239000007758 minimum essential medium Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 235000021049 nutrient content Nutrition 0.000 description 1
- 210000000287 oocyte Anatomy 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000008024 pharmaceutical diluent Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 229920000036 polyvinylpyrrolidone Chemical class 0.000 description 1
- 239000001267 polyvinylpyrrolidone Chemical class 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 230000001177 retroviral effect Effects 0.000 description 1
- 239000002336 ribonucleotide Substances 0.000 description 1
- 125000002652 ribonucleotide group Chemical group 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 239000012090 serum-supplement Substances 0.000 description 1
- 230000020509 sex determination Effects 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 230000004096 skeletal muscle tissue growth Effects 0.000 description 1
- 235000015424 sodium Nutrition 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 235000010413 sodium alginate Nutrition 0.000 description 1
- 239000000661 sodium alginate Substances 0.000 description 1
- 229940005550 sodium alginate Drugs 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241000701447 unidentified baculovirus Species 0.000 description 1
- 210000003501 vero cell Anatomy 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 229940075420 xanthine Drugs 0.000 description 1
- 210000001325 yolk sac Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/111—General methods applicable to biologically active non-coding nucleic acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
- C12N15/1136—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against growth factors, growth regulators, cytokines, lymphokines or hormones
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
- A01K2217/054—Animals comprising random inserted nucleic acids (transgenic) inducing loss of function
- A01K2217/058—Animals comprising random inserted nucleic acids (transgenic) inducing loss of function due to expression of inhibitory nucleic acid, e.g. siRNA, antisense
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/30—Bird
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/02—Animal zootechnically ameliorated
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2320/00—Applications; Uses
- C12N2320/30—Special therapeutic applications
- C12N2320/32—Special delivery means, e.g. tissue-specific
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Endocrinology (AREA)
- Veterinary Medicine (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
配列番号1-ニワトリミオスタチン(Genbank NM_001001461)。
配列番号2-ニワトリミオスタチンをコードするヌクレオチド配列(Genbank NM_001001461)。
配列番号3-ニワトリDMRT1の部分タンパク質配列(Genbank AF123456)。
配列番号4-ニワトリDMRT1をコードする部分核酸配列(Genbank AF123456)。
配列番号5-ニワトリWPKCI(ASW)(Genbank AF148455)。
配列番号6-ニワトリWPKCI(ASW)をコードするヌクレオチド配列(Genbank AF148455)。
配列番号7-ニワトリU6-1プロモーターのヌクレオチド配列。
配列番号8-ニワトリU6-3プロモーターのヌクレオチド配列。
配列番号9-ニワトリU6-4プロモーターのヌクレオチド配列。
配列番号10-ニワトリ7SKプロモーターのヌクレオチド配列。
配列番号11〜98および113〜122-本発明に有用なRNA配列。
配列番号99〜112-オリゴヌクレオチドプライマー。
別段に具体的に定義しない限り、本明細書で用いられる全ての技術的および科学用語は、(例えば、細胞培養、分子遺伝学、トリの生物学、RNA干渉および生化学において)当業者に通常理解されるのと同じ意味を有すると解されるべきである。
用語「RNA干渉」、「RNAi」または「遺伝子発現抑制」は、全般的に、2本鎖RNA(dsRNA)分子が実質的にまたは完全に相同性を有する核酸配列の発現を、該2本鎖RNA分子が低減させるプロセスのことをいう。しかし、遺伝子発現抑制は、非RNA 2本鎖分子を用いて達成できることがより最近になって示されている(例えば米国特許出願公開第20070004667号を参照されたい)。
本発明の方法は、トリの種の任意の形質、特に卵内での胚の発生中に決定または影響される形質を改変するために用いることができる。改変され得る好ましい形質は、性別および筋肉量を含む。
CCAGUUGUCAAGAAGAGCA(配列番号11)
GGAUGCUCAUUCAGGACAU(配列番号12)
CCCUGUAUCCUUACUAUAA(配列番号13)
GCCACUGAGUCUUCCUCAA(配列番号14)
CCAGCAACAUACAUGUCAA(配列番号15)
CCUGCGUCACACAGAUACU(配列番号16)
GGAGUAGUUGUACAGGUUG(配列番号17)
GACUGGCUUGACAUGUAUG(配列番号18)
AUGGCGGUUCUCCAUCCCU(配列番号19)
AAGCUAGCAGUCUAUGUUU(配列番号20)
GCUAGCAGUCUAUGUUUAU(配列番号21)
CGCUGAAAAAGACGGACUG(配列番号22)
AAAGACGGACUGUGCAAUG(配列番号23)
AGACGGACUGUGCAAUGCU(配列番号24)
UGCUUGUACGUGGAGACAG(配列番号25)
UACAAAAUCCUCCAGAAUA(配列番号26)
AAUCCUCCAGAAUAGAAGC(配列番号27)
UCCUCCAGAAUAGAAGCCA(配列番号28)
UAGAAGCCAUAAAAAUUCA(配列番号29)
GCCAUAAAAAUUCAAAUCC(配列番号30)
AAAUUCAAAUCCUCAGCAA(配列番号31)
AUUCAAAUCCUCAGCAAAC(配列番号32)
AUCCUCAGCAAACUGCGCC(配列番号33)
ACUGCGCCUGGAACAAGCA(配列番号34)
CAAGCACCUAACAUUAGCA(配列番号35)
GCACCUAACAUUAGCAGGG(配列番号36)
CAUUAGCAGGGACGUUAUU(配列番号37)
GCAGCUUUUACCCAAAGCU(配列番号38)
UUCCUGCAGUGGAGGAGCU(配列番号39)
CUGAUUGAUCAGUAUGAUG(配列番号40)
GACGAUGACUAUCAUGCCA(配列番号41)
CCGAGACGAUUAUCACAAU(配列番号42)
UGCCUACGGAGUCUGAUUU(配列番号43)
AUGGAGGGAAAACCAAAAU(配列番号44)
AACCAAAAUGUUGCUUCUU(配列番号45)
CCAAAAUGUUGCUUCUUUA(配列番号46)
AAUGUUGCUUCUUUAAGUU(配列番号47)
UGUUGCUUCUUUAAGUUUA(配列番号48)
GUUUAGCUCUAAAAUACAA(配列番号49)
AAUACAAUAUAACAAAGUA(配列番号50)
UACAAUAUAACAAAGUAGU(配列番号51)
UAUAACAAAGUAGUAAAGG(配列番号52)
CAAAGUAGUAAAGGCACAA(配列番号53)
AGUAGUAAAGGCACAAUUA(配列番号54)
AGGCACAAUUAUGGAUAUA(配列番号55)
UUAUGGAUAUACUUGAGGC(配列番号56)
GUCCAAAAACCUACAACGG(配列番号57)
AAACCUACAACGGUGUUUG(配列番号58)
ACCUACAACGGUGUUUGUG(配列番号59)
CGGUGUUUGUGCAGAUCCU(配列番号60)
GCCCAUGAAAGACGGUACA(配列番号61)
AGACGGUACAAGAUAUACU(配列番号62)
GAUAUACUGGAAUUCGAUC(配列番号63)
UUCGAUCUUUGAAACUUGA(配列番号64)
ACUUGACAUGAACCCAGGC(配列番号65)
CCCAGGCACUGGUAUCUGG(配列番号66)
GACAGUGCUGCAAAAUUGG(配列番号67)
AAUUGGCUCAAACAGCCUG(配列番号68)
UUGGCUCAAACAGCCUGAA(配列番号69)
ACAGCCUGAAUCCAAUUUA(配列番号70)
UCCAAUUUAGGCAUCGAAA(配列番号71)
UUUAGGCAUCGAAAUAAAA(配列番号72)
AUAAAAGCUUUUGAUGAGA(配列番号73)
AAGCUUUUGAUGAGACUGG(配列番号74)
GCUUUUGAUGAGACUGGAC(配列番号75)
GAUGGAUUGAACCCAUUUU(配列番号76)
CCCAUUUUUAGAGGUCAGA(配列番号77)
ACGGUCCCGCAGAGAUUUU(配列番号78)
CGGAAUCCCGAUGUUGUCG(配列番号79)
UCCAGUCCCAUCCAAAAGC(配列番号80)
GCUUUUGGAUGGGACUGGA(配列番号81)
AAGAUACAAAGCCAAUUAC(配列番号82)
GAUACAAAGCCAAUUACUG(配列番号83)
AGCCAAUUACUGCUCCGGA(配列番号84)
UUACUGCUCCGGAGAAUGC(配列番号85)
UGCGAAUUUGUGUUUCUAC(配列番号86)
CAGGUGAGUGUGCGGGUAU(配列番号87)
AUACCCGCACACUCACCUG(配列番号88)
GCAAAUCCCAGAGGUCCAG(配列番号89)
AUCCCAGAGGUCCAGCAGG(配列番号90)
GAUGUCCCCUAUAAACAUG(配列番号91)
ACAUGCUGUAUUUCAAUGG(配列番号92)
UGGAAAAGAACAAAUAAUA(配列番号93)
AAGAACAAAUAAUAUAUGG(配列番号94)
GAACAAAUAAUAUAUGGAA(配列番号95)
CAAAUAAUAUAUGGAAAGA(配列番号96)
AUAAUAUAUGGAAAGAUAC(配列番号97)
UAUAUGGAAAGAUACCAGC(配列番号98)
CCAGAAUAGAAGCCAUAAA(配列番号113)
GCACAAUUAUGGAUAUACU(配列番号114)
GUACAAGAUAUACUGGAAU(配列番号115)
CCUAUAAACAUGCUGUAUU(配列番号116)
GCGAAUUUGUGUUUCUACA(配列番号117)
GAGUAUUGAUGUGAAGACA(配列番号118)
CCUCCAGAAUAGAAGCCAU(配列番号119)
GGUCAGAGUUACAGACACA(配列番号120)
CAGUGGAUUUCGAAGCUUU(配列番号121)
CAACGGUGUUUGUGCAGAU(配列番号122)
本発明は、本発明の2本鎖領域またはその1本鎖を含む核酸分子をコードするベクターも提供する。好ましくは、ベクターは、宿主細胞および/または無細胞系にてdsRNAをコードするオープンリーディングフレーム(複数可)を発現できる発現ベクターである。宿主細胞は、それらに限定されないが細菌、真菌、植物または動物の細胞のような任意の型の細胞であり得る。
本発明は、トリの卵に投与できる2本鎖領域を含む核酸分子を含む組成物も提供する。2本鎖領域を含む核酸分子を含む組成物は、組成物を投与に適したものにするために、医薬的に許容される担体を含有してよい。
2本鎖領域を含む核酸分子(2本鎖領域を含む核酸分子を含む組成物を含む)の投与は、卵への注入、一般的には気嚢への注入により、簡便に達成される。in ovo投与の好ましい経路は気嚢であるが、卵黄嚢または卵膜尿膜液のようなその他の領域へ注入により接種してもよい。孵化率は、投与の標的が気嚢でない場合はわずかに低減するが、必ずしも商業的に受け入れがたいレベルではない。針が卵または発生中の胚もしくは胚を取り囲む胚体外膜の組織および器官に過度の損傷を引き起こさないことが好ましいが、注入の機構は本発明の実行に重要でない。
(実施例1)
ニワトリでのDMRT1タンパク質産生を下方制御するためのshRNA分子の同定
DMRT1を標的にするshRNA配列の選択
本発明者らは、Ambionにより設計されたsiRNAターゲットファインダー(www.ambion.com/techlib/misc/siRNA_finder.html)を用いて、Dmrt1についての30個の予測siRNA配列を選択した。30個のsiRNA配列を、次いで、shRNAの選択のためにスクリーニングした(Table1(表1))。特定の標的遺伝子についての可能性のあるsiRNA配列を選択するために用い得るいくつかのアルゴリズムが存在する。しかし、これらの予測siRNAの多くが、発現されたshRNAからプロセシングされたときに有効に機能しないことが示されている。Taxmanら(2006)は、有効なshRNA分子を予測するアルゴリズムを特に設計し、本発明者らは、shRNA予測を改良するためにそのアルゴリズムに独自の改変を加えた。本発明者らは、Dmrt1遺伝子発現の特異的ノックダウンのためのshRNAとして試験するための配列を選択するように、改変Taxmanアルゴリズムを、30個の選択されたsiRNAに適用した。
ニワトリポリメラーゼIIIプロモーターcU6-1(GenBankアクセッション番号DQ531567)およびcU6-4(DQ531570)を鋳型として用いて、選択したdmrt1および対照(EGFPおよび無関係の)shRNAのためのddRNAi発現プラスミドを、1ステップPCRにより構築した(図1)。shRNAプラスミドの構築のためのPCRは、プライマーTD175とTH346(shDmrt1-346用)、TH461(shDmrt1-461)、TH566(shDmrt1-566)、TH622(shDmrt1-622)、TH697(shDmrt1-697)、TH839(shDmrt1-839)またはTD195(shEGFP)との対を用いた(プライマー配列および増幅された具体的なshRNAの詳細についてTable 3(表3)を参照されたい)。各PCRでのリバースプライマーは、各プロモーター配列の最後の20nt、shRNAセンス、ループおよびshRNAアンチセンス配列を含むように設計し、HPLCで精製した。全長の増幅した発現カセット産物をpGEM-T Easyに連結し、次いで配列決定した。遺伝子ノックダウンアッセイに用いた最終的なshRNA発現プラスミドは、pshDmrt1-346、pshDmrt1-461、pshDmrt1-566、pshDmrt1-622、pshDmrt1-697、pshDmrt1-839およびpshEGFPと命名した。cU6-1の無関係の対照プラスミドも構築し、遺伝子発現アッセイにおいて模擬比較として用いた(以下を参照されたい)。この模擬プラスミドについて、フォワードプライマーTD135を、chU6-1プロモーターの最後の20ntおよびその他の全ての無関係のshRNA成分を含むリバースプライマーTD149と対にした。PCR産物を、pGEM-T Easyに連結し、配列決定した。
レポーター遺伝子発現アッセイを用いて、shRNAを、Dmrt1の発現抑制について試験した。レポーター遺伝子は、pEGFP-C(Clontech)中の高感度緑色蛍光タンパク質(EGFP)遺伝子の3'末端の下流に挿入されたDmrt1の転写遺伝子融合体であった。レポータープラスミドは、次のようにして構築した。Dmrt1のcDNAを、4日齢の胚から単離した全RNAから逆転写し、pCMV-Script(Stratagene)のマルチクローニング部位にクローニングした。Dmrt1挿入断片を、NotI-EcoRI断片としてクローニングベクターから回収し、pEGFP-C(Clontech)中のEGFP遺伝子の下流にクローニングした。得られたプラスミドは、pEGFP-Dmrt1と命名した。このプラスミドを、ニワトリDF-1細胞にトランスフェクションし、転写遺伝子融合体の発現を、以下に記載するようにフローサイトメトリーを用いてEGFP蛍光を測定することにより確認した。DF-1細胞は、ニワトリ胚繊維芽細胞の継続系統で、EV-0胚(ATCC、CRL-12203)に由来するので、RNAi分子のin ovo効果を研究するためのモデル系である。
ニワトリでのミオスタチンタンパク質産生を下方制御するためのshRNA分子の同定
ミオスタチン(Gdf8)を標的にするshRNA配列の選択
79個のGdf8についての予測siRNA配列を、Ambionにより設計されたsiRNAターゲットファインダー(www.ambion.com/techlib/misc/siRNA_finder.html)を用いて同定した(Table4(表4))。Taxmanアルゴリズムを用いて最適化した付加的なsiRNA配列を、Table5(表5)に示す。本発明者らは、これらの配列のうち3つ(Gdf8-258、Gdf8-913およびGdf8-1002)を、shRNAの発現のためのddRNAiプラスミドの構築のために選択した(Table4(表4)において太字で示す)。
ニワトリポリメラーゼIIIプロモーターcU6-1(GenBankアクセッション番号DQ531567)を鋳型として用いて、選択したGdf8およびcEGFP shRNAのためのddRNAi発現プラスミドを、1ステップPCRにより構築した(図1)。shRNAプラスミドの構築のためのPCRは、プライマーTD 135とDS304(shGdf8-253用)、DS305(shGdf8-913)、DS306(shGdf8-1002)またはTD148(shEGFP)との対を用いた(プライマー配列および増幅された具体的なshRNAについてTable6(表6)を参照されたい)。各PCRでのリバースプライマーは、各プロモーター配列の最後の20nt、shRNAセンス、ループおよびshRNAアンチセンス配列を含むように設計し、HPLCで精製した。全長の増幅した発現カセット産物をpGEM-T Easyに連結し、次いで配列決定した。遺伝子ノックダウンアッセイに用いた最終的なshRNA発現プラスミドは、pshGdf8-253、pshGdf8-913、pshGdf8-1002およびpshEGFPと命名した。
レポーター遺伝子発現アッセイを用いて、3つの選択したshRNAを、Gdf8の発現抑制について試験した。レポーター遺伝子は、pEGFP-C(Clontech)中の高感度緑色蛍光タンパク質(EGFP)の3'末端の下流に挿入されたGdf8の転写遺伝子融合体であった。レポータープラスミドは、次のようにして構築した。Gdf8のcDNAを、7日齢の胚から単離した全RNAから逆転写し、pGEM-T Easy(Promega)のマルチクローニング部位にクローニングした。Gdf8挿入断片を、NotI断片としてクローニングベクターから回収し、pEGFP-C(Clontech)中のEGFP遺伝子の下流にクローニングした。得られたプラスミドは、pEGFP-Gdf8と命名した。このプラスミドを、ニワトリDF-1細胞にトランスフェクションし、転写遺伝子融合体の発現を、以下に記載するようにフローサイトメトリーを用いてEGFP蛍光を測定することにより確認した。
Amarzguioui et al. (2004) Biochem Biophys Res Commun 316:1050-1058
Elbashire et al. (2001) Nature 411 :494-498 Hori et al. (2000) MoI Biol Cell 11 :3645-3660
Freier et al. (1986) Proc Natl Acad Sci USA 83:9373-9377
Needleman and Wunsch (1970) J MoI Biol 48: 443-453
O'Neill et al. (2000) Dev Genes Evol 210:243-249
Raymond et al. (1999) Dev Biol. 215:208-220 Reynolds et al. (2004) Nat. Biotech., 22:326-330
Smith et al. (1999) Nature 402:601-602
Smith et al. (2000) Nature 407: 319-320.
Taxman et al. (2006) BMC Biotechnol, Jan 24, 6:7
Waterhouse et al. (1998) Proc Natl Acad Sci USA 95:13959-13964
Claims (23)
- トリの形質を改変する方法であって、トリの卵に2本鎖領域を含む少なくとも1つの核酸分子を投与する段階を含み、前記核酸分子が前記卵内の少なくとも1つのRNA分子および/またはタンパク質のレベルの低減をもたらす方法。
- 前記核酸分子がdsRNAである、請求項1に記載の方法。
- 前記dsRNAがsiRNAまたはshRNAである、請求項2に記載の方法。
- 前記形質が生産形質である、請求項1から3のいずれか一項に記載の方法。
- 前記生産形質が筋肉量または性別である、請求項4に記載の方法。
- 前記生産形質が性別であり、前記核酸分子がDMRT1遺伝子によりコードされるタンパク質のレベルを低減させる、請求項5に記載の方法。
- 前記核酸分子が、
CCAGUUGUCAAGAAGAGCA(配列番号11)
GGAUGCUCAUUCAGGACAU(配列番号12)
CCCUGUAUCCUUACUAUAA(配列番号13)
GCCACUGAGUCUUCCUCAA(配列番号14)
CCAGCAACAUACAUGUCAA(配列番号15)
CCUGCGUCACACAGAUACU(配列番号16)
GGAGUAGUUGUACAGGUUG(配列番号17)
GACUGGCUUGACAUGUAUG(配列番号18)
AUGGCGGUUCUCCAUCCCU(配列番号19)、
またはそれらのいずれか1つの変異体から選択される少なくとも1つのヌクレオチド配列を含む、請求項6に記載の方法。 - 前記生産形質が筋肉量であり、前記核酸分子がミオスタチン遺伝子によりコードされるタンパク質のレベルを低減させる、請求項5に記載の方法。
- 前記核酸分子が、配列CAGGUGAGUGUGCGGGUAU(配列番号87)またはその変異体を含む、請求項8に記載の方法。
- 前記核酸分子が注入により投与される、請求項1から9のいずれか一項に記載の方法。
- 前記トリが、ニワトリ、カモ、シチメンチョウ、ガチョウ、チャボおよびウズラから選択される、請求項1から10のいずれか一項に記載の方法。
- 請求項1から11のいずれか一項に記載の方法を用いて生産されたトリ。
- 請求項1から11のいずれか一項に記載の方法を用いて生産されたニワトリ。
- トリの卵に投与されたときに少なくとも1つのRNA分子および/またはタンパク質のレベルを低減させる2本鎖領域を含む単離および/または外因性核酸分子。
- dsRNA分子である、請求項14に記載の核酸分子。
- DMRT1遺伝子またはミオスタチン遺伝子によりコードされるタンパク質のレベルを低減させる、請求項14または請求項15に記載の核酸分子。
- CCAGUUGUCAAGAAGAGCA(配列番号11)
GGAUGCUCAUUCAGGACAU(配列番号12)
CCCUGUAUCCUUACUAUAA(配列番号13)
GCCACUGAGUCUUCCUCAA(配列番号14)
CCAGCAACAUACAUGUCAA(配列番号15)
CCUGCGUCACACAGAUACU(配列番号16)
GGAGUAGUUGUACAGGUUG(配列番号17)
GACUGGCUUGACAUGUAUG(配列番号18)
AUGGCGGUUCUCCAUCCCU(配列番号19)
AAGCUAGCAGUCUAUGUUU(配列番号20)
GCUAGCAGUCUAUGUUUAU(配列番号21)
CGCUGAAAAAGACGGACUG(配列番号22)
AAAGACGGACUGUGCAAUG(配列番号23)
AGACGGACUGUGCAAUGCU(配列番号24)
UGCUUGUACGUGGAGACAG(配列番号25)
UACAAAAUCCUCCAGAAUA(配列番号26)
AAUCCUCCAGAAUAGAAGC(配列番号27)
UCCUCCAGAAUAGAAGCCA(配列番号28)
UAGAAGCCAUAAAAAUUCA(配列番号29)
GCCAUAAAAAUUCAAAUCC(配列番号30)
AAAUUCAAAUCCUCAGCAA(配列番号31)
AUUCAAAUCCUCAGCAAAC(配列番号32)
AUCCUCAGCAAACUGCGCC(配列番号33)
ACUGCGCCUGGAACAAGCA(配列番号34)
CAAGCACCUAACAUUAGCA(配列番号35)
GCACCUAACAUUAGCAGGG(配列番号36)
CAUUAGCAGGGACGUUAUU(配列番号37)
GCAGCUUUUACCCAAAGCU(配列番号38)
UUCCUGCAGUGGAGGAGCU(配列番号39)
CUGAUUGAUCAGUAUGAUG(配列番号40)
GACGAUGACUAUCAUGCCA(配列番号41)
CCGAGACGAUUAUCACAAU(配列番号42)
UGCCUACGGAGUCUGAUUU(配列番号43)
AUGGAGGGAAAACCAAAAU(配列番号44)
AACCAAAAUGUUGCUUCUU(配列番号45)
CCAAAAUGUUGCUUCUUUA(配列番号46)
AAUGUUGCUUCUUUAAGUU(配列番号47)
UGUUGCUUCUUUAAGUUUA(配列番号48)
GUUUAGCUCUAAAAUACAA(配列番号49)
AAUACAAUAUAACAAAGUA(配列番号50)
UACAAUAUAACAAAGUAGU(配列番号51)
UAUAACAAAGUAGUAAAGG(配列番号52)
CAAAGUAGUAAAGGCACAA(配列番号53)
AGUAGUAAAGGCACAAUUA(配列番号54)
AGGCACAAUUAUGGAUAUA(配列番号55)
UUAUGGAUAUACUUGAGGC(配列番号56)
GUCCAAAAACCUACAACGG(配列番号57)
AAACCUACAACGGUGUUUG(配列番号58)
ACCUACAACGGUGUUUGUG(配列番号59)
CGGUGUUUGUGCAGAUCCU(配列番号60)
GCCCAUGAAAGACGGUACA(配列番号61)
AGACGGUACAAGAUAUACU(配列番号62)
GAUAUACUGGAAUUCGAUC(配列番号63)
UUCGAUCUUUGAAACUUGA(配列番号64)
ACUUGACAUGAACCCAGGC(配列番号65)
CCCAGGCACUGGUAUCUGG(配列番号66)
GACAGUGCUGCAAAAUUGG(配列番号67)
AAUUGGCUCAAACAGCCUG(配列番号68)
UUGGCUCAAACAGCCUGAA(配列番号69)
ACAGCCUGAAUCCAAUUUA(配列番号70)
UCCAAUUUAGGCAUCGAAA(配列番号71)
UUUAGGCAUCGAAAUAAAA(配列番号72)
AUAAAAGCUUUUGAUGAGA(配列番号73)
AAGCUUUUGAUGAGACUGG(配列番号74)
GCUUUUGAUGAGACUGGAC(配列番号75)
GAUGGAUUGAACCCAUUUU(配列番号76)
CCCAUUUUUAGAGGUCAGA(配列番号77)
ACGGUCCCGCAGAGAUUUU(配列番号78)
CGGAAUCCCGAUGUUGUCG(配列番号79)
UCCAGUCCCAUCCAAAAGC(配列番号80)
GCUUUUGGAUGGGACUGGA(配列番号81)
AAGAUACAAAGCCAAUUAC(配列番号82)
GAUACAAAGCCAAUUACUG(配列番号83)
AGCCAAUUACUGCUCCGGA(配列番号84)
UUACUGCUCCGGAGAAUGC(配列番号85)
UGCGAAUUUGUGUUUCUAC(配列番号86)
CAGGUGAGUGUGCGGGUAU(配列番号87)
AUACCCGCACACUCACCUG(配列番号88)
GCAAAUCCCAGAGGUCCAG(配列番号89)
AUCCCAGAGGUCCAGCAGG(配列番号90)
GAUGUCCCCUAUAAACAUG(配列番号91)
ACAUGCUGUAUUUCAAUGG(配列番号92)
UGGAAAAGAACAAAUAAUA(配列番号93)
AAGAACAAAUAAUAUAUGG(配列番号94)
GAACAAAUAAUAUAUGGAA(配列番号95)
CAAAUAAUAUAUGGAAAGA(配列番号96)
AUAAUAUAUGGAAAGAUAC(配列番号97)
UAUAUGGAAAGAUACCAGC(配列番号98)
CCAGAAUAGAAGCCAUAAA(配列番号113)
GCACAAUUAUGGAUAUACU(配列番号114)
GUACAAGAUAUACUGGAAU(配列番号115)
CCUAUAAACAUGCUGUAUU(配列番号116)
GCGAAUUUGUGUUUCUACA(配列番号117)
GAGUAUUGAUGUGAAGACA(配列番号118)
CCUCCAGAAUAGAAGCCAU(配列番号119)
GGUCAGAGUUACAGACACA(配列番号120)
CAGUGGAUUUCGAAGCUUU(配列番号121)
CAACGGUGUUUGUGCAGAU(配列番号122)、
またはそれらのいずれか1つの変異体から選択される少なくとも1つのヌクレオチド配列を含む、請求項16に記載の核酸分子。 - 請求項14から17のいずれか一項に記載の核酸分子またはその1本鎖をコードするベクター。
- 請求項14から17のいずれか一項に記載の外因性核酸分子もしくはその1本鎖および/または請求項18に記載のベクターを含む宿主細胞。
- 請求項14から17のいずれか一項に記載の核酸分子もしくはその1本鎖、請求項18に記載のベクターおよび/または請求項19に記載の宿主細胞を含む組成物。
- 請求項14から17のいずれか一項に記載の核酸分子もしくはその1本鎖、請求項18に記載のベクターおよび/または請求項19に記載の宿主細胞を含むトリの卵。
- 請求項14から17のいずれか一項に記載の核酸分子もしくはその1本鎖、請求項18に記載のベクター、請求項19に記載の宿主細胞および/または請求項20に記載の組成物を含むキット。
- 個々にまたはまとめて特許請求の範囲に開示され、または本出願の明細書に示される段階、特徴、整数、組成物および/または化合物、ならびに前記段階または特徴の2つ以上のありとあらゆる組合せ。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US94370807P | 2007-06-13 | 2007-06-13 | |
US60/943,708 | 2007-06-13 | ||
PCT/AU2008/000835 WO2008151364A1 (en) | 2007-06-13 | 2008-06-12 | Modulating production traits in avians |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2010528667A true JP2010528667A (ja) | 2010-08-26 |
JP5554231B2 JP5554231B2 (ja) | 2014-07-23 |
Family
ID=40129126
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2010511446A Expired - Fee Related JP5554231B2 (ja) | 2007-06-13 | 2008-06-12 | トリの生産形質の改変 |
Country Status (9)
Country | Link |
---|---|
US (2) | US20100306869A1 (ja) |
EP (1) | EP2167645A4 (ja) |
JP (1) | JP5554231B2 (ja) |
CN (1) | CN101802173B (ja) |
AU (1) | AU2008261604B2 (ja) |
BR (1) | BRPI0812500A2 (ja) |
MX (1) | MX2009013532A (ja) |
RU (1) | RU2518681C2 (ja) |
WO (1) | WO2008151364A1 (ja) |
Families Citing this family (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102356158B (zh) * | 2008-12-17 | 2014-04-02 | 联邦科学技术研究组织 | 调节禽类性别的方法 |
AU2010210315A1 (en) * | 2009-02-08 | 2011-08-18 | Murdoch Childrens Research Institute | Sex-determination and methods of specifying same |
CN102041257B (zh) * | 2009-10-13 | 2013-05-22 | 中国农业大学 | 抑制鸡肌肉生成抑制素基因表达的siRNA及其应用 |
WO2012177639A2 (en) * | 2011-06-22 | 2012-12-27 | Alnylam Pharmaceuticals, Inc. | Bioprocessing and bioproduction using avian cell lines |
US11172656B2 (en) | 2012-12-11 | 2021-11-16 | Signify Holding B.V. | Methods for controlling sex of oviparous embryos using light sources |
US10455819B2 (en) | 2012-12-11 | 2019-10-29 | Signify North America Corporation | Methods for controlling sex of oviparous embryos using light sources |
US11140879B2 (en) | 2012-12-11 | 2021-10-12 | Signify North America Corporation | Methods for controlling sex of oviparous embryos using light sources |
CN104918482B (zh) | 2012-12-11 | 2019-02-01 | 万斯创新公司 | 使用光控制卵生胚胎的性别 |
US10004814B2 (en) * | 2013-11-11 | 2018-06-26 | Sirna Therapeutics, Inc. | Systemic delivery of myostatin short interfering nucleic acids (siNA) conjugated to a lipophilic moiety |
US11051495B2 (en) | 2015-09-15 | 2021-07-06 | Signify North America Corporation | Systems and methods for promoting biological responses in incubated eggs |
US10933081B2 (en) | 2016-09-21 | 2021-03-02 | Alnylam Pharmaceuticals, Inc. | Myostatin iRNA compositions and methods of use thereof |
CA3065317A1 (en) * | 2017-05-31 | 2018-12-06 | Commonwealth Scientific And Industrial Research Organisation | Trait selection in avians |
CN110791569B (zh) * | 2018-08-03 | 2022-05-10 | 浙江省农业科学院 | 一种鸭蛋蛋壳颜色相关分子标记及其应用 |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH07504320A (ja) * | 1992-01-27 | 1995-05-18 | ノース・カロライナ・ステート・ユニバーシティ | インオボ(in ovo)で筋肉内にDNAを導入することによる鳥類への遺伝子伝達 |
JP2002534066A (ja) * | 1999-01-06 | 2002-10-15 | エンブレクス,インコーポレイテッド | 卵内への注射と検出を同時に行う方法及びその装置 |
US20030068821A1 (en) * | 2001-09-13 | 2003-04-10 | Carlos Lois-Caballe | Method for expression of small RNA molecules within a cell |
JP2004517612A (ja) * | 2000-11-02 | 2004-06-17 | アクゾ・ノベル・エヌ・ベー | マーカーワクチンとしての、組み換えニューカッスル病ウイルス核タンパク質の突然変異体 |
JP2004535194A (ja) * | 2001-06-14 | 2004-11-25 | ザ ユニバーシティ オブ メルボルン | 弱毒化サーコウイルス |
JP2007512027A (ja) * | 2003-11-21 | 2007-05-17 | レビビコア, インコーポレイテッド | トランスジェニック動物の産生における干渉rnaの使用 |
Family Cites Families (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4417663A (en) * | 1981-06-16 | 1983-11-29 | Kiyonobu Suzuki | Apparatus for determining the sex of a chick |
US4903635A (en) * | 1986-07-02 | 1990-02-27 | Embrex, Inc. | High speed automated injection system for avian embryos |
US5056464A (en) * | 1990-01-18 | 1991-10-15 | Embrex, Inc. | Automated injection system for avian embryos with advanced fluid delivery system |
US5508165A (en) * | 1990-09-21 | 1996-04-16 | Zoogen, Inc. | Avian sex determination probe |
US5136979A (en) * | 1991-09-25 | 1992-08-11 | Embrex, Inc. | Modular injection system for avian embryos |
JPH07206705A (ja) * | 1993-11-03 | 1995-08-08 | American Cyanamid Co | 生インオボ(in ovo)ワクチン |
WO1999051755A2 (en) * | 1998-04-03 | 1999-10-14 | The Salk Institute For Biological Studies | Ribozyme-mediated control of gene expression |
US6130365A (en) * | 1998-08-03 | 2000-10-10 | Peterson Farms | Breeding lines of color-sexable day-old chicks and methods for producing the same |
US8202979B2 (en) * | 2002-02-20 | 2012-06-19 | Sirna Therapeutics, Inc. | RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid |
US7662791B2 (en) * | 2000-08-02 | 2010-02-16 | University Of Southern California | Gene silencing using mRNA-cDNA hybrids |
AUPR064600A0 (en) * | 2000-10-09 | 2000-11-02 | Commonwealth Scientific And Industrial Research Organisation | Genetic control of sex ratio in animal populations |
RU2189743C1 (ru) * | 2001-01-30 | 2002-09-27 | Орловский государственный аграрный университет | Способ регуляции пола у животных, например, крупного рогатого скота |
ES2401326T3 (es) * | 2001-11-21 | 2013-04-18 | Astellas Pharma Inc. | Procedimiento para inhibir la expresión génica |
US7617795B2 (en) * | 2004-10-13 | 2009-11-17 | Embrex, Inc. | Methods and apparatus for injecting and sampling material through avian egg membranes |
US20060196428A1 (en) * | 2005-03-03 | 2006-09-07 | Correa Rafael S | Method for improving chick hatchability |
CN102356158B (zh) * | 2008-12-17 | 2014-04-02 | 联邦科学技术研究组织 | 调节禽类性别的方法 |
-
2008
- 2008-06-12 JP JP2010511446A patent/JP5554231B2/ja not_active Expired - Fee Related
- 2008-06-12 CN CN200880102770.XA patent/CN101802173B/zh not_active Expired - Fee Related
- 2008-06-12 EP EP08756916A patent/EP2167645A4/en not_active Ceased
- 2008-06-12 AU AU2008261604A patent/AU2008261604B2/en not_active Ceased
- 2008-06-12 BR BRPI0812500A patent/BRPI0812500A2/pt not_active IP Right Cessation
- 2008-06-12 RU RU2010100886/10A patent/RU2518681C2/ru not_active IP Right Cessation
- 2008-06-12 US US12/664,097 patent/US20100306869A1/en not_active Abandoned
- 2008-06-12 WO PCT/AU2008/000835 patent/WO2008151364A1/en active Application Filing
- 2008-06-12 MX MX2009013532A patent/MX2009013532A/es active IP Right Grant
-
2012
- 2012-09-04 US US13/602,927 patent/US20130067606A1/en not_active Abandoned
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH07504320A (ja) * | 1992-01-27 | 1995-05-18 | ノース・カロライナ・ステート・ユニバーシティ | インオボ(in ovo)で筋肉内にDNAを導入することによる鳥類への遺伝子伝達 |
JP2002534066A (ja) * | 1999-01-06 | 2002-10-15 | エンブレクス,インコーポレイテッド | 卵内への注射と検出を同時に行う方法及びその装置 |
JP2004517612A (ja) * | 2000-11-02 | 2004-06-17 | アクゾ・ノベル・エヌ・ベー | マーカーワクチンとしての、組み換えニューカッスル病ウイルス核タンパク質の突然変異体 |
JP2004535194A (ja) * | 2001-06-14 | 2004-11-25 | ザ ユニバーシティ オブ メルボルン | 弱毒化サーコウイルス |
US20030068821A1 (en) * | 2001-09-13 | 2003-04-10 | Carlos Lois-Caballe | Method for expression of small RNA molecules within a cell |
JP2007512027A (ja) * | 2003-11-21 | 2007-05-17 | レビビコア, インコーポレイテッド | トランスジェニック動物の産生における干渉rnaの使用 |
Non-Patent Citations (8)
Title |
---|
JPN6013024855; Developmental Dynamics Vol.231, 2004, p.592-600 * |
JPN6013024856; Developmental Biology Vol.285, 2005, p.80-90 * |
JPN6013024858; Biotechnology & Genetic Enginerring Reviews Vol.22, 2006, p.63-75 * |
JPN6013024859; Oligonucleotides Vol.13, No.5, 2003, p.411-419 * |
JPN6013024861; Journal of RNAi and Gene Silencing Vol.2, No.1, 2006, p.126-135 * |
JPN6013024862; Nature Biotechnology Vol.21, 2003, p.93-96 * |
JPN6013024864; 生物工学会誌 Vol.83, No.9, 2005, p.450 * |
JPN6013024866; BioEssays Vol.26, 2004, p.120-132 * |
Also Published As
Publication number | Publication date |
---|---|
RU2518681C2 (ru) | 2014-06-10 |
EP2167645A1 (en) | 2010-03-31 |
US20100306869A1 (en) | 2010-12-02 |
MX2009013532A (es) | 2010-03-04 |
RU2010100886A (ru) | 2011-07-20 |
WO2008151364A1 (en) | 2008-12-18 |
CN101802173B (zh) | 2014-09-24 |
US20130067606A1 (en) | 2013-03-14 |
BRPI0812500A2 (pt) | 2019-09-24 |
JP5554231B2 (ja) | 2014-07-23 |
AU2008261604A1 (en) | 2008-12-18 |
EP2167645A4 (en) | 2012-04-18 |
AU2008261604B2 (en) | 2012-05-24 |
CN101802173A (zh) | 2010-08-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5554231B2 (ja) | トリの生産形質の改変 | |
AU2009328633B2 (en) | Methods of modulating the sex of avians | |
Westenberg et al. | siRNA injection induces sequence-independent protection in Penaeus monodon against white spot syndrome virus | |
US7589189B2 (en) | Inhibition of the expression of huntingtin gene | |
AU2008250973B2 (en) | Treatment and prevention of influenza | |
WO2017088017A1 (en) | Production of viruses in avian eggs | |
BR112019025358A2 (pt) | ovo aviário transgênico, ave transgênica, ovo ou progênie aviária, método para detectar um ovo aviário macho, método para produzir um ovo aviário, método para replicar um vírus, vírus produzidos pelo uso de ovo aviário, método para produzir uma composição de vacina, composição da vacina produzida pelo uso do método e método para produzir um ovo aviário transgênico ou uma ave produzida pelo ovo | |
WO2018084077A1 (ja) | 有用エビ類の卵成熟抑制を解除する方法 | |
US20140289881A1 (en) | Double-stranded rna | |
Derrick et al. | Lamb1a regulates atrial growth by limiting second heart field addition during zebrafish heart development | |
AU2012204092B2 (en) | Modulating production traits in avians | |
WO2014153590A1 (en) | Rna interference in amoebas | |
AU2013202277A1 (en) | Methods of modulating the sex of avians | |
JP4877835B2 (ja) | Rna干渉誘導エレメント及びその用途 | |
US20050037962A1 (en) | Syndecans and angiogenesis | |
JP2022521463A (ja) | 新規魚トティウイルス | |
AU2013200109A1 (en) | Treatment and prevention of influenza | |
KR20110093840A (ko) | 지방세포 형성의 조절을 위한 plac8 활성 억제제의 용도 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20110608 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20120803 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20120803 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20130528 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20130822 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20130822 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20140428 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20140528 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 5554231 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
LAPS | Cancellation because of no payment of annual fees |