JPWO2019189827A1 - 核酸構築物、医薬組成物、抗がん剤、抗ウイルス剤及び抗菌剤 - Google Patents
核酸構築物、医薬組成物、抗がん剤、抗ウイルス剤及び抗菌剤 Download PDFInfo
- Publication number
- JPWO2019189827A1 JPWO2019189827A1 JP2020511128A JP2020511128A JPWO2019189827A1 JP WO2019189827 A1 JPWO2019189827 A1 JP WO2019189827A1 JP 2020511128 A JP2020511128 A JP 2020511128A JP 2020511128 A JP2020511128 A JP 2020511128A JP WO2019189827 A1 JPWO2019189827 A1 JP WO2019189827A1
- Authority
- JP
- Japan
- Prior art keywords
- rna
- nucleic acid
- virus
- acid construct
- cleaving
- 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
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 88
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 80
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 80
- 239000003242 anti bacterial agent Substances 0.000 title claims description 10
- 239000003443 antiviral agent Substances 0.000 title claims description 10
- 239000002246 antineoplastic agent Substances 0.000 title claims description 8
- 239000008194 pharmaceutical composition Substances 0.000 title claims description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims abstract description 85
- 108020005004 Guide RNA Proteins 0.000 claims abstract description 57
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 55
- 241000700605 Viruses Species 0.000 claims abstract description 38
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 37
- 241000894006 Bacteria Species 0.000 claims abstract description 21
- 241000251539 Vertebrata <Metazoa> Species 0.000 claims abstract description 21
- 230000014509 gene expression Effects 0.000 claims abstract description 20
- 230000035772 mutation Effects 0.000 claims abstract description 19
- 102000040650 (ribonucleotides)n+m Human genes 0.000 claims abstract description 11
- 210000004027 cell Anatomy 0.000 claims description 70
- 108020004414 DNA Proteins 0.000 claims description 23
- 230000005945 translocation Effects 0.000 claims description 16
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 8
- 239000004480 active ingredient Substances 0.000 claims description 8
- 241000712461 unidentified influenza virus Species 0.000 claims description 5
- 208000009746 Adult T-Cell Leukemia-Lymphoma Diseases 0.000 claims description 3
- 208000016683 Adult T-cell leukemia/lymphoma Diseases 0.000 claims description 3
- 241000725619 Dengue virus Species 0.000 claims description 3
- 241001115402 Ebolavirus Species 0.000 claims description 3
- 241000991587 Enterovirus C Species 0.000 claims description 3
- 208000007514 Herpes zoster Diseases 0.000 claims description 3
- 241000725303 Human immunodeficiency virus Species 0.000 claims description 3
- 241000712079 Measles morbillivirus Species 0.000 claims description 3
- 241000127282 Middle East respiratory syndrome-related coronavirus Species 0.000 claims description 3
- 241000711386 Mumps virus Species 0.000 claims description 3
- 241001263478 Norovirus Species 0.000 claims description 3
- 241001631646 Papillomaviridae Species 0.000 claims description 3
- 241000702670 Rotavirus Species 0.000 claims description 3
- 241000315672 SARS coronavirus Species 0.000 claims description 3
- 201000006966 adult T-cell leukemia Diseases 0.000 claims description 3
- 208000006454 hepatitis Diseases 0.000 claims description 3
- 231100000283 hepatitis Toxicity 0.000 claims description 3
- 241001529453 unidentified herpesvirus Species 0.000 claims description 3
- 208000010201 Exanthema Diseases 0.000 claims description 2
- 208000014770 Foot disease Diseases 0.000 claims description 2
- 201000005884 exanthem Diseases 0.000 claims description 2
- 206010037844 rash Diseases 0.000 claims description 2
- 206010028980 Neoplasm Diseases 0.000 description 22
- 201000011510 cancer Diseases 0.000 description 18
- 239000013612 plasmid Substances 0.000 description 11
- 230000000694 effects Effects 0.000 description 10
- 230000004927 fusion Effects 0.000 description 10
- 108091027544 Subgenomic mRNA Proteins 0.000 description 8
- 206010042863 synovial sarcoma Diseases 0.000 description 8
- 238000007702 DNA assembly Methods 0.000 description 7
- 238000010362 genome editing Methods 0.000 description 7
- 210000000805 cytoplasm Anatomy 0.000 description 6
- 230000008685 targeting Effects 0.000 description 6
- 101710137189 Amyloid-beta A4 protein Proteins 0.000 description 5
- 101710151993 Amyloid-beta precursor protein Proteins 0.000 description 5
- 102100022704 Amyloid-beta precursor protein Human genes 0.000 description 5
- 208000003174 Brain Neoplasms Diseases 0.000 description 5
- 108090001102 Hammerhead ribozyme Proteins 0.000 description 5
- 108091035715 XIST (gene) Proteins 0.000 description 5
- 108091033409 CRISPR Proteins 0.000 description 4
- 208000035473 Communicable disease Diseases 0.000 description 4
- 102100035102 E3 ubiquitin-protein ligase MYCBP2 Human genes 0.000 description 4
- 238000000636 Northern blotting Methods 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 238000003776 cleavage reaction Methods 0.000 description 4
- 238000010276 construction Methods 0.000 description 4
- 210000001808 exosome Anatomy 0.000 description 4
- 108020004999 messenger RNA Proteins 0.000 description 4
- 239000013642 negative control Substances 0.000 description 4
- 230000007017 scission Effects 0.000 description 4
- 102000012758 APOBEC-1 Deaminase Human genes 0.000 description 3
- 108010079649 APOBEC-1 Deaminase Proteins 0.000 description 3
- 102100033311 APOBEC1 complementation factor Human genes 0.000 description 3
- 101000799953 Homo sapiens APOBEC1 complementation factor Proteins 0.000 description 3
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 3
- 238000010357 RNA editing Methods 0.000 description 3
- 230000026279 RNA modification Effects 0.000 description 3
- 108091007416 X-inactive specific transcript Proteins 0.000 description 3
- 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 Chemical class 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 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- -1 inhalants Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 239000013603 viral vector Substances 0.000 description 3
- 208000024827 Alzheimer disease Diseases 0.000 description 2
- 102000002659 Amyloid Precursor Protein Secretases Human genes 0.000 description 2
- 108010043324 Amyloid Precursor Protein Secretases Proteins 0.000 description 2
- 102000013455 Amyloid beta-Peptides Human genes 0.000 description 2
- 108010090849 Amyloid beta-Peptides Proteins 0.000 description 2
- 241000272517 Anseriformes Species 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 241000282472 Canis lupus familiaris Species 0.000 description 2
- 108090000994 Catalytic RNA Proteins 0.000 description 2
- 102000053642 Catalytic RNA Human genes 0.000 description 2
- 108010077544 Chromatin Proteins 0.000 description 2
- 108091026890 Coding region Proteins 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
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000282326 Felis catus Species 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 2
- 206010064571 Gene mutation Diseases 0.000 description 2
- 241000029603 Leptotrichia shahii Species 0.000 description 2
- 241000711408 Murine respirovirus Species 0.000 description 2
- 241000588652 Neisseria gonorrhoeae Species 0.000 description 2
- 230000007022 RNA scission Effects 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 241000607768 Shigella Species 0.000 description 2
- 108091027967 Small hairpin RNA Proteins 0.000 description 2
- 108020004459 Small interfering RNA Proteins 0.000 description 2
- 241000251131 Sphyrna Species 0.000 description 2
- 241000282887 Suidae Species 0.000 description 2
- 241000607626 Vibrio cholerae Species 0.000 description 2
- DZHSAHHDTRWUTF-SIQRNXPUSA-N amyloid-beta polypeptide 42 Chemical compound C([C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O)[C@@H](C)CC)C(C)C)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C(C)C)C1=CC=CC=C1 DZHSAHHDTRWUTF-SIQRNXPUSA-N 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 235000013330 chicken meat Nutrition 0.000 description 2
- 210000003483 chromatin Anatomy 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 239000000645 desinfectant Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 210000001320 hippocampus Anatomy 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 108091027963 non-coding RNA Proteins 0.000 description 2
- 102000042567 non-coding RNA Human genes 0.000 description 2
- 108091092562 ribozyme Proteins 0.000 description 2
- 239000004055 small Interfering RNA Substances 0.000 description 2
- 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 2
- 239000013598 vector Substances 0.000 description 2
- 229940118696 vibrio cholerae Drugs 0.000 description 2
- 241000589291 Acinetobacter Species 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 241000588832 Bordetella pertussis Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000589562 Brucella Species 0.000 description 1
- 108091079001 CRISPR RNA Proteins 0.000 description 1
- 238000010354 CRISPR gene editing Methods 0.000 description 1
- 238000010356 CRISPR-Cas9 genome editing Methods 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 241000589876 Campylobacter Species 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 241001647372 Chlamydia pneumoniae Species 0.000 description 1
- 241000588923 Citrobacter Species 0.000 description 1
- 241000702421 Dependoparvovirus Species 0.000 description 1
- 208000004232 Enteritis Diseases 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 108091092584 GDNA Proteins 0.000 description 1
- 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 1
- 241000590002 Helicobacter pylori Species 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 102000006947 Histones Human genes 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000880769 Homo sapiens Protein SSX1 Proteins 0.000 description 1
- 101000880775 Homo sapiens Protein SSX5 Proteins 0.000 description 1
- 101000759186 Homo sapiens Zinc finger translocation-associated protein Proteins 0.000 description 1
- 241000588748 Klebsiella Species 0.000 description 1
- 241000589242 Legionella pneumophila Species 0.000 description 1
- 241000029590 Leptotrichia wadei Species 0.000 description 1
- 241000186779 Listeria monocytogenes Species 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000204031 Mycoplasma Species 0.000 description 1
- 108010077850 Nuclear Localization Signals Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 241000282579 Pan Species 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- 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 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 102100037687 Protein SSX1 Human genes 0.000 description 1
- 102100037723 Protein SSX5 Human genes 0.000 description 1
- 241000588769 Proteus <enterobacteria> Species 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 230000014632 RNA localization Effects 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 208000006265 Renal cell carcinoma Diseases 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 241000607598 Vibrio Species 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 210000001766 X chromosome Anatomy 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 102100023386 Zinc finger translocation-associated protein Human genes 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000012298 atmosphere Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 239000012459 cleaning agent Substances 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000001416 effect on chromosomes Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 208000010932 epithelial neoplasm Diseases 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000003889 eye drop Substances 0.000 description 1
- 229940012356 eye drops Drugs 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940037467 helicobacter pylori Drugs 0.000 description 1
- 230000002008 hemorrhagic effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 229940115932 legionella pneumophila Drugs 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 230000002101 lytic effect Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000001394 metastastic effect Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 229940127084 other anti-cancer agent Drugs 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000004853 protein function Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 231100000164 trypan blue assay Toxicity 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Images
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
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/22—Ribonucleases RNAses, DNAses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
-
- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
-
- 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
-
- 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
- C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPRs]
-
- 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
- C12N2800/00—Nucleic acids vectors
- C12N2800/80—Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Epidemiology (AREA)
- Virology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Description
項1. 1又は複数の標的RNAに結合する少なくとも1種のガイドRNA部分とRNA切断型Casタンパク質発現部分を含み、前記標的RNAは脊椎動物細胞の突然変異、ウイルス又は細菌に由来するものである、核酸構築物。
項2. 前記ガイドRNA部分と前記RNA切断型Casタンパク質発現部分が1つの核酸配列中に存在する、項1に記載の核酸構築物。
項3. 前記ガイドRNA部分と前記RNA切断型Casタンパク質発現部分が別の核酸配列中に存在し、2以上の核酸を含む、項1に記載の核酸構築物。
項4. 核酸構築物がRNA構築物又はDNA構築物である、項1〜3のいずれか1項に記載の核酸構築物。
項5. RNA切断型Casタンパク質がCas13ファミリータンパク質である、項1〜4のいずれか1項に記載の核酸構築物。
項6. RNA切断型Casタンパク質がC2C2/Cas13aである、項1〜5のいずれか1項に記載の核酸構築物。
項7. ガイドRNAが脊椎動物細胞の突然変異に対応するRNAを標的とする、項1〜6のいずれか1項に記載の核酸構築物。
項8. 脊椎動物細胞の突然変異が転座であり、ガイドRNAは転座遺伝子に対応するRNAを標的とする、項1〜7のいずれか1項に記載の核酸構築物。
項9. 前記ウイルスがインフルエンザウイルス、HIVウイルス、ヘルペスウイルス、エボラウイルス、トリインフルエンザウイルス、口蹄疫ウイルス、SARSコロナウイルス、MERSコロナウイルス、パピローマウイルス、肝炎ウイルス(A型、B型、C型)、麻疹ウイルス、風疹ウイルス、ムンプスウイルス、ロタウイルス、RSウイルス、ノロウイルス、帯状疱疹ウイルス、ポリオウイルス、デングウイルス及び成人T細胞白血病ウイルスからなる群から選ばれるいずれかである、項1〜6のいずれか1項に記載の核酸構築物。
項10. 項1〜8のいずれか1項に記載の核酸構築物を有効成分とする、医薬組成物。
項11. 項1〜8のいずれか1項に記載の核酸構築物を有効成分とする、抗がん剤。
項12. 項1〜8のいずれか1項に記載の核酸構築物を有効成分とする、抗ウイルス剤。
項13. 項1〜8のいずれか1項に記載の核酸構築物を有効成分とする、抗菌剤。
実施例1
(1)プラスミドの構築と標的ガイド
C2C2 Lsh(Leptotrichia shahii)DNA配列を増幅し、Hifi DNA Assembly (NEB)を用いてpX458プラスミドに融合した。Hifi DNA Assembly (NEB)を用いてLsh特異的scaffold RNA配列を挿入した。sgRNA配列をコードするリン酸化オリゴヌクレオチドをBbs1-消化scaffold構築物にライゲートしてsgRNA標的化XistRNA, C11orf95-RELA融合RNA及びSS18-SSX 融合RNAを調製した。このプラットフォームをpLMTと名付けた(pLMT_Xistプラスミド)。15種のpLMT_Xistプラスミドは、表1に示す15種のガイドRNAのいずれかを各々含む。
HEK293TとSYO-1滑膜肉腫細胞株を、1%ペニシリンとストレプトマイシン及び10%ウシ胎児血清(FBS)を添加したD-MEM (低グルコース)培地で維持した。得られた細胞を加湿雰囲気下に5% CO2、37℃で培養した。
(3)ノーザンブロッティング
ScreenFect A (Wako)を用いてpLMT_Xistプラスミドを30%コンフルエンシーのHEK293T細胞にトランスフェクトした。10種のpLMT_Xistプラスミドは、表1に示す上皮腫 h.C11orf95RELA fusion Guide RNA listのいずれかのガイドRNAを含むものである。48時間培養後、細胞を収穫した。ISOGENを用いてRNAを沈殿させた。DIG Northern Starter kit (Roche)を用いてノーザンブロッティングを行った。メンブランをハイブリダイゼーションバッファー(7% SDS, 0.5M Na-phosphste buffer (pH 7.2), 10mM EDTA)中でXistRNAとC11-orf95-RELA を標的化したDIG-標識プローブとハイブリダイズし、洗浄バッファー(1% SDS, Na-phosphste buffer (pH 7.2), 10mM EDTA)で洗浄した。ノーザンブロッティングの結果を図4に示す。
(4)トリパンブルーアッセイ
ScreenFect A (Wako)を用い、pLMT_Xistプラスミドを30%コンフルエンシーのSYO-1細胞にトランスフェクトした。48時間培養後、細胞を収穫した。細胞懸濁液と0.4%トリパンブルー溶液と1:1で混合した。生細胞及び死細胞を血球計算盤でカウントし、死細胞の割合を測定した。結果を図2に示す。
(1)プラスミドの構築と標的ガイド
C2C2 Lsh(Leptotrichia shahii)DNA配列を増幅し、Hifi DNA Assembly (NEB)を用いてpX458プラスミドに融合した。Hifi DNA Assembly (NEB)を用いてLsh特異的scaffold RNA配列を挿入した。sgRNA配列をコードするリン酸化オリゴヌクレオチドをBbs1-消化scaffold構築物にライゲートしてsgRNA標的化XistRNA,融合RNAを調製した。RNA高次構造予測にしたがってデザインされ、HEK293Tに導入された71種のガイドRNA配列を、以下の表2〜表4に示す。また、図5に導入された71種のガイドRNAとその実験条件と結果を示す。
(1)プラスミドの構築と標的ガイド
PAマグネットシステム作成のため、Dead Lwa(別名dCas13a: Leptotrichia wadei由来)DNA配列を二分割増幅し、Hifi DNA Assembly (NEB)を用いてpcDNA3.1-PAに融合した(これをpPA-dCas13-EGFPと呼ぶ)。Hifi DNA Assembly (NEB)を用いてLwa特異的scaffold RNA配列を挿入した。sgRNA配列をコードするリン酸化オリゴヌクレオチドをBbs1-消化scaffold構築物にライゲートしてsgRNA標的化XistRNA,融合RNAを調製した。
(2)標的RNA局在確認
RNA高次構造予測にしたがってデザインされた短鎖配列標的sgRNAとpPA-dCas13-EGFPをHEK293T細胞に導入した。標的となるXIST RNAへの結合を蛍光顕微鏡にて確認した(図6)。
(3)標的RNA機能評価 ChIP-qPCR
標的となるXIST RNAのクロマチン上における任意配列への結合を確認した。RNA高次構造予測にしたがってデザインされた短鎖配列標的sgRNAとpPA-dCas13-EGFPをHEK293T細胞に導入したのちに、1%パラフォルムアルデヒドにてクロマチン上にあるXIST-sgRNA-Cas13-EGFP複合体を架橋させ、細胞抽出液内にあるCas13-EGFP融合タンパク質を抗GFP抗体を用いて免疫沈降させた後に共同免疫沈降されたXISTとXIST結合ゲノム配列の抽出を行った。抽出されたゲノム配列はqPCR法にて検出を行い、XIST-クロマチンの重合を確認した。陽性対照としてヒストン、陰性対象として非特異的IgGを用いている(図7)。
(1)プラスミドの構築と標的ガイド
RNA書き換えシステム作成のため、Hifi DNA Assembly (NEB)を用いてpPA-dCas13-EGFPのEGFPドメインをAPOBEC1ドメインおよびA1CFドメイン融合ドメインに書き換えた(RESCUE system: pPA-dCas13-ABC1A1と呼ぶ。図8)。sgRNA配列をコードするリン酸化オリゴヌクレオチドをBbs1-消化scaffold構築物にライゲートしてsgRNA標的化APPタンパク質のAβ切断配列認識RNA融合RNAを調製した。
(2)標的RNA遺伝子書き換え効果検証
標的APPタンパク質の機能とRNA高次構造予測にしたがってデザインされた短鎖配列標的sgRNAとpPA-dCas13-ABC1A1をHEK293T細胞に導入した。標的となるAPPタンパク質のβセクレターゼによる切断抑制効果をウェスタンブロット解析にて確認した(図9)。
Claims (13)
- 1又は複数の標的RNAに結合する少なくとも1種のガイドRNA部分とRNA切断型Casタンパク質発現部分を含み、前記標的RNAは脊椎動物細胞の突然変異、ウイルス又は細菌に由来するものである、核酸構築物。
- 前記ガイドRNA部分と前記RNA切断型Casタンパク質発現部分が1つの核酸配列中に存在する、請求項1に記載の核酸構築物。
- 前記ガイドRNA部分と前記RNA切断型Casタンパク質発現部分が別の核酸配列中に存在し、2以上の核酸を含む、請求項1に記載の核酸構築物。
- 核酸構築物がRNA構築物又はDNA構築物である、請求項1〜3のいずれか1項に記載の核酸構築物。
- RNA切断型Casタンパク質がCas13ファミリータンパク質である、請求項1〜4のいずれか1項に記載の核酸構築物。
- RNA切断型Casタンパク質がC2C2/Cas13aである、請求項1〜5のいずれか1項に記載の核酸構築物。
- ガイドRNAが脊椎動物細胞の突然変異に対応するRNAを標的とする、請求項1〜6のいずれか1項に記載の核酸構築物。
- 脊椎動物細胞の突然変異が転座であり、ガイドRNAは転座遺伝子に対応するRNAを標的とする、請求項1〜7のいずれか1項に記載の核酸構築物。
- 前記ウイルスがインフルエンザウイルス、HIVウイルス、ヘルペスウイルス、エボラウイルス、トリインフルエンザウイルス、口蹄疫ウイルス、SARSコロナウイルス、MERSコロナウイルス、パピローマウイルス、肝炎ウイルス(A型、B型、C型)、麻疹ウイルス、風疹ウイルス、ムンプスウイルス、ロタウイルス、RSウイルス、ノロウイルス、帯状疱疹ウイルス、ポリオウイルス、デングウイルス及び成人T細胞白血病ウイルスからなる群から選ばれるいずれかである、請求項1〜6のいずれか1項に記載の核酸構築物。
- 請求項1〜8のいずれか1項に記載の核酸構築物を有効成分とする、医薬組成物。
- 請求項1〜8のいずれか1項に記載の核酸構築物を有効成分とする、抗がん剤。
- 請求項1〜8のいずれか1項に記載の核酸構築物を有効成分とする、抗ウイルス剤。
- 請求項1〜8のいずれか1項に記載の核酸構築物を有効成分とする、抗菌剤。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2018067565 | 2018-03-30 | ||
JP2018067565 | 2018-03-30 | ||
PCT/JP2019/014183 WO2019189827A1 (ja) | 2018-03-30 | 2019-03-29 | 核酸構築物、医薬組成物、抗がん剤、抗ウイルス剤及び抗菌剤 |
Publications (2)
Publication Number | Publication Date |
---|---|
JPWO2019189827A1 true JPWO2019189827A1 (ja) | 2021-04-08 |
JP7113395B2 JP7113395B2 (ja) | 2022-08-05 |
Family
ID=68061992
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2020511128A Active JP7113395B2 (ja) | 2018-03-30 | 2019-03-29 | 核酸構築物、医薬組成物、抗がん剤、抗ウイルス剤及び抗菌剤 |
Country Status (3)
Country | Link |
---|---|
US (1) | US20210189386A1 (ja) |
JP (1) | JP7113395B2 (ja) |
WO (1) | WO2019189827A1 (ja) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113337488B (zh) * | 2020-03-02 | 2024-04-19 | 中国科学院分子细胞科学卓越创新中心 | 一种分离的Cas13蛋白 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016205764A1 (en) * | 2015-06-18 | 2016-12-22 | The Broad Institute Inc. | Novel crispr enzymes and systems |
WO2017219027A1 (en) * | 2016-06-17 | 2017-12-21 | The Broad Institute Inc. | Type vi crispr orthologs and systems |
-
2019
- 2019-03-29 JP JP2020511128A patent/JP7113395B2/ja active Active
- 2019-03-29 WO PCT/JP2019/014183 patent/WO2019189827A1/ja active Application Filing
- 2019-03-29 US US17/043,138 patent/US20210189386A1/en active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016205764A1 (en) * | 2015-06-18 | 2016-12-22 | The Broad Institute Inc. | Novel crispr enzymes and systems |
WO2017219027A1 (en) * | 2016-06-17 | 2017-12-21 | The Broad Institute Inc. | Type vi crispr orthologs and systems |
Non-Patent Citations (11)
Title |
---|
ABUDAYYEH, OMAR O. ET AL.: "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector", SCIENCE, vol. 353, JPN6019043723, 2016, pages 5537 - 1, ISSN: 0004567579 * |
ABUDAYYEH, OMAR O. ET AL.: "RNA targeting with CRISPR-Cas13", NATURE, vol. 550, JPN6021007073, 2017, pages 280 - 284, XP055529736, ISSN: 0004567572, DOI: 10.1038/nature24049 * |
ACUNZO, MARIO ET AL.: "Selective targeting of point-mutated KRAS through artificial microRNAs", PROC. NATL. ACAD. SCI. U. S. A., vol. 114, JPN6021030621, 2017, pages 4203 - 4212, ISSN: 0004567575 * |
AMAN, RASHID ET AL.: "RNA virus interference via CRISPR/Cas13a system in plants", GENOME BIOL., vol. Vol. 19: 1, JPN6019022914, 4 January 2018 (2018-01-04), pages 1 - 9, XP055639663, ISSN: 0004567578, DOI: 10.1186/s13059-017-1381-1 * |
COX, DAVID B. T. ET AL.: "RNA editing with CRISPR-Cas13", SCIENCE, vol. 358, JPN6019022909, 2017, pages 1019 - 1027, XP055491658, ISSN: 0004567576, DOI: 10.1126/science.aaq0180 * |
KONERMANN, SILVANA ET AL.: "Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors", CELL, vol. 173, JPN6019022912, 2018, pages 665 - 676, XP055529705, ISSN: 0004567577, DOI: 10.1016/j.cell.2018.02.033 * |
NIHONGAKI, YUTA ET AL.: "Photoactivatable CRISPR-Cas9 for optogenetic genome editing", NAT. BIOTECHNOL., vol. 33, JPN6019043727, 2015, pages 755 - 760, XP055538990, ISSN: 0004684954, DOI: 10.1038/nbt.3245 * |
SU, LE ET AL.: "Deconstruction of the SS18-SSX fusion oncoprotein complex: insights into disease etiology and therap", CANCER CELL, vol. 21, JPN6020006780, 2012, pages 333 - 347, XP028473636, ISSN: 0004453853, DOI: 10.1016/j.ccr.2012.01.010 * |
TERNS, MICHAEL P.: "CRISPR-Based Technologies: Impact of RNA-Targeting Systems", MOL. CELL, vol. 72, JPN6021030620, 1 November 2018 (2018-11-01), pages 404 - 412, XP085522306, ISSN: 0004684955, DOI: 10.1016/j.molcel.2018.09.018 * |
THOMAS, MARIA ET AL.: "Targeting leukemic fusion proteins with small interfering RNAs: recent advances and therapeutic pote", ACTA PHARMACOL. SIN., vol. 27, JPN6020006779, 2006, pages 273 - 281, ISSN: 0004684952 * |
ZAREE MAHMODABADY, ALI ET AL.: "Bcr-abl silencing by specific small-interference RNA expression vector as a potential treatment for", IRAN. BIOMED. J., vol. 14, JPN6020006781, 2010, pages 1 - 8, ISSN: 0004567580 * |
Also Published As
Publication number | Publication date |
---|---|
US20210189386A1 (en) | 2021-06-24 |
WO2019189827A1 (ja) | 2019-10-03 |
JP7113395B2 (ja) | 2022-08-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US11667903B2 (en) | Tracking and manipulating cellular RNA via nuclear delivery of CRISPR/CAS9 | |
US10689691B2 (en) | Unbiased identification of double-strand breaks and genomic rearrangement by genome-wide insert capture sequencing | |
Shioda et al. | Targeting G-quadruplex DNA as cognitive function therapy for ATR-X syndrome | |
Lee et al. | Nanoparticle delivery of CRISPR into the brain rescues a mouse model of fragile X syndrome from exaggerated repetitive behaviours | |
Zuris et al. | Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo | |
JP6914274B2 (ja) | Crisprcpf1の結晶構造 | |
US11001829B2 (en) | Functional screening with optimized functional CRISPR-Cas systems | |
US10377998B2 (en) | CRISPR-CAS systems and methods for altering expression of gene products, structural information and inducible modular CAS enzymes | |
Wang et al. | Transformation of the intestinal epithelium by the MSI2 RNA-binding protein | |
US11492670B2 (en) | Compositions and methods for targeting cancer-specific sequence variations | |
EP3647421A1 (en) | Using minivectors to treat ovarian cancer | |
JP7389135B2 (ja) | タウ凝集に関連する遺伝的脆弱性を明らかにするためのcrispr/casドロップアウトスクリーニングプラットフォーム | |
El-Sawy et al. | Nickel stimulates L1 retrotransposition by a post-transcriptional mechanism | |
JP2024071489A (ja) | タウの播種または凝集の遺伝的修飾因子を同定するためのcrispr/casスクリーニングプラットフォーム | |
WO2023241669A1 (zh) | CRISPR-Cas效应子蛋白、其基因编辑系统及应用 | |
JP7113395B2 (ja) | 核酸構築物、医薬組成物、抗がん剤、抗ウイルス剤及び抗菌剤 | |
JP2021523733A (ja) | 動物病原体由来ポリペプチドおよびその遺伝子工学的利用 | |
Ghasemi et al. | Interstrand crosslinking of homologous repair template DNA enhances gene editing in human cells | |
US20080045471A1 (en) | Nucleic Acids For Apoptosis Of Cancer Cells | |
JP7212943B2 (ja) | がん遺伝子の転写調節領域 | |
Krachulec et al. | Footprintless disruption of prosurvival genes in aneuploid cancer cells using CRISPR/Cas9 technology | |
CN111989113B (zh) | 用于治疗癌症的包含向导rna和核酸内切酶作活性成分的药物组合物 | |
Yang et al. | Expanded CAG/CTG repeats resist gene silencing mediated by targeted epigenome editing | |
EP4347815A2 (en) | Compositions and methods for increasing efficiency of precise editing repair | |
Gahan | The biology of CNAPS |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A529 | Written submission of copy of amendment under article 34 pct |
Free format text: JAPANESE INTERMEDIATE CODE: A5211 Effective date: 20200907 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20200914 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20201125 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20201125 |
|
A871 | Explanation of circumstances concerning accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A871 Effective date: 20201125 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20201207 |
|
A975 | Report on accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A971005 Effective date: 20210208 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20210302 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20210427 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20210810 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20211001 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20211124 |
|
A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20220118 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20220413 |
|
C60 | Trial request (containing other claim documents, opposition documents) |
Free format text: JAPANESE INTERMEDIATE CODE: C60 Effective date: 20220413 |
|
A911 | Transfer to examiner for re-examination before appeal (zenchi) |
Free format text: JAPANESE INTERMEDIATE CODE: A911 Effective date: 20220420 |
|
C21 | Notice of transfer of a case for reconsideration by examiners before appeal proceedings |
Free format text: JAPANESE INTERMEDIATE CODE: C21 Effective date: 20220426 |
|
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: 20220524 |
|
A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20220615 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20220615 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20220615 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 7113395 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |