JP4118579B2 - Ecat5ノックアウト細胞 - Google Patents
Ecat5ノックアウト細胞 Download PDFInfo
- Publication number
- JP4118579B2 JP4118579B2 JP2002073138A JP2002073138A JP4118579B2 JP 4118579 B2 JP4118579 B2 JP 4118579B2 JP 2002073138 A JP2002073138 A JP 2002073138A JP 2002073138 A JP2002073138 A JP 2002073138A JP 4118579 B2 JP4118579 B2 JP 4118579B2
- Authority
- JP
- Japan
- Prior art keywords
- ecat5
- cells
- gene
- seq
- knockout
- 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.)
- Expired - Fee Related
Links
- 108090000623 proteins and genes Proteins 0.000 claims description 110
- 206010043276 Teratoma Diseases 0.000 claims description 14
- 230000015572 biosynthetic process Effects 0.000 claims description 7
- 241000124008 Mammalia Species 0.000 claims description 3
- 108091026890 Coding region Proteins 0.000 claims 5
- 210000004027 cell Anatomy 0.000 description 166
- 239000013598 vector Substances 0.000 description 35
- 230000014509 gene expression Effects 0.000 description 25
- 239000002299 complementary DNA Substances 0.000 description 20
- 239000013604 expression vector Substances 0.000 description 20
- 241000699666 Mus <mouse, genus> Species 0.000 description 19
- 241000699670 Mus sp. Species 0.000 description 17
- 238000000034 method Methods 0.000 description 16
- 108020004414 DNA Proteins 0.000 description 14
- 239000012634 fragment Substances 0.000 description 14
- 238000011813 knockout mouse model Methods 0.000 description 14
- 108010014186 ras Proteins Proteins 0.000 description 14
- 102000016914 ras Proteins Human genes 0.000 description 14
- 238000010367 cloning Methods 0.000 description 13
- 239000002953 phosphate buffered saline Substances 0.000 description 13
- 238000004458 analytical method Methods 0.000 description 12
- 238000002105 Southern blotting Methods 0.000 description 11
- 239000002609 medium Substances 0.000 description 11
- 108091006027 G proteins Proteins 0.000 description 10
- 102000030782 GTP binding Human genes 0.000 description 10
- 108091000058 GTP-Binding Proteins 0.000 description 10
- 102000004058 Leukemia inhibitory factor Human genes 0.000 description 10
- 108090000581 Leukemia inhibitory factor Proteins 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 210000004962 mammalian cell Anatomy 0.000 description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 9
- 210000004436 artificial bacterial chromosome Anatomy 0.000 description 9
- 239000000872 buffer Substances 0.000 description 9
- 230000004069 differentiation Effects 0.000 description 9
- 108010010803 Gelatin Proteins 0.000 description 8
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 8
- 235000001014 amino acid Nutrition 0.000 description 8
- 229940024606 amino acid Drugs 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 8
- 239000000499 gel Substances 0.000 description 8
- 239000008273 gelatin Substances 0.000 description 8
- 229920000159 gelatin Polymers 0.000 description 8
- 235000019322 gelatine Nutrition 0.000 description 8
- 235000011852 gelatine desserts Nutrition 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 238000001262 western blot Methods 0.000 description 8
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 7
- 230000030570 cellular localization Effects 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 7
- 238000002649 immunization Methods 0.000 description 7
- 230000003053 immunization Effects 0.000 description 7
- 239000012528 membrane Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 7
- 230000035755 proliferation Effects 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 230000008685 targeting Effects 0.000 description 7
- 238000005406 washing Methods 0.000 description 7
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 6
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 6
- 150000001413 amino acids Chemical group 0.000 description 6
- 239000012636 effector Substances 0.000 description 6
- 238000004520 electroporation Methods 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 238000010363 gene targeting Methods 0.000 description 6
- 238000002744 homologous recombination Methods 0.000 description 6
- 230000006801 homologous recombination Effects 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 108091060211 Expressed sequence tag Proteins 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 5
- 206010028980 Neoplasm Diseases 0.000 description 5
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 5
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000009396 hybridization Methods 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 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 4
- 241000588724 Escherichia coli Species 0.000 description 4
- 101710113436 GTPase KRas Proteins 0.000 description 4
- HVLSXIKZNLPZJJ-TXZCQADKSA-N HA peptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HVLSXIKZNLPZJJ-TXZCQADKSA-N 0.000 description 4
- 108091034117 Oligonucleotide Proteins 0.000 description 4
- 108090000631 Trypsin Proteins 0.000 description 4
- 102000004142 Trypsin Human genes 0.000 description 4
- 239000011543 agarose gel Substances 0.000 description 4
- 239000004202 carbamide Substances 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 230000036961 partial effect Effects 0.000 description 4
- 230000037361 pathway Effects 0.000 description 4
- 230000006798 recombination Effects 0.000 description 4
- 238000005215 recombination Methods 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 239000012588 trypsin Substances 0.000 description 4
- 239000011534 wash buffer Substances 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 description 3
- 102000013446 GTP Phosphohydrolases Human genes 0.000 description 3
- 102100039788 GTPase NRas Human genes 0.000 description 3
- 108091006109 GTPases Proteins 0.000 description 3
- 101000744505 Homo sapiens GTPase NRas Proteins 0.000 description 3
- 241000829388 Mus musculus polyomavirus 1 Species 0.000 description 3
- 238000000636 Northern blotting Methods 0.000 description 3
- 102000003993 Phosphatidylinositol 3-kinases Human genes 0.000 description 3
- 108090000430 Phosphatidylinositol 3-kinases Proteins 0.000 description 3
- 102000000113 Ral guanine nucleotide dissociation stimulator Human genes 0.000 description 3
- 108050008437 Ral guanine nucleotide dissociation stimulator Proteins 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 210000001766 X chromosome Anatomy 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 208000031513 cyst Diseases 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 108020001507 fusion proteins Proteins 0.000 description 3
- 102000037865 fusion proteins Human genes 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000035772 mutation Effects 0.000 description 3
- 239000002773 nucleotide Substances 0.000 description 3
- 125000003729 nucleotide group Chemical group 0.000 description 3
- 230000004481 post-translational protein modification Effects 0.000 description 3
- 230000002062 proliferating effect Effects 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- 210000001082 somatic cell Anatomy 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 229920000936 Agarose Polymers 0.000 description 2
- 241000972773 Aulopiformes Species 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 108010005939 Ciliary Neurotrophic Factor Proteins 0.000 description 2
- 102100031614 Ciliary neurotrophic factor Human genes 0.000 description 2
- 108700024394 Exon Proteins 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 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
- 101710128836 Large T antigen Proteins 0.000 description 2
- 102100021747 Leukemia inhibitory factor receptor Human genes 0.000 description 2
- 101710142062 Leukemia inhibitory factor receptor Proteins 0.000 description 2
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- 102000004140 Oncostatin M Human genes 0.000 description 2
- 108090000630 Oncostatin M Proteins 0.000 description 2
- 101150099493 STAT3 gene Proteins 0.000 description 2
- 235000010724 Wisteria floribunda Nutrition 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- -1 aliphatic amino acid Chemical class 0.000 description 2
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- 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 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- 210000001367 artery Anatomy 0.000 description 2
- 210000002459 blastocyst Anatomy 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- YTRQFSDWAXHJCC-UHFFFAOYSA-N chloroform;phenol Chemical compound ClC(Cl)Cl.OC1=CC=CC=C1 YTRQFSDWAXHJCC-UHFFFAOYSA-N 0.000 description 2
- 210000004748 cultured cell Anatomy 0.000 description 2
- 235000018417 cysteine Nutrition 0.000 description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 230000013020 embryo development Effects 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 239000003797 essential amino acid Substances 0.000 description 2
- 235000020776 essential amino acid Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 230000016784 immunoglobulin production Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 231100000518 lethal Toxicity 0.000 description 2
- 230000001665 lethal effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000013011 mating Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000009456 molecular mechanism Effects 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 229930002330 retinoic acid Natural products 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 235000019515 salmon Nutrition 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 229960001727 tretinoin Drugs 0.000 description 2
- 230000005740 tumor formation Effects 0.000 description 2
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 210000004291 uterus Anatomy 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- 102000006822 Agouti Signaling Protein Human genes 0.000 description 1
- 108010072151 Agouti Signaling Protein Proteins 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 101000583086 Bunodosoma granuliferum Delta-actitoxin-Bgr2b Proteins 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 241000484025 Cuniculus Species 0.000 description 1
- 108700003861 Dominant Genes Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 239000012591 Dulbecco’s Phosphate Buffered Saline Substances 0.000 description 1
- 108010067770 Endopeptidase K Proteins 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
- 241000282326 Felis catus Species 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101001059454 Homo sapiens Serine/threonine-protein kinase MARK2 Proteins 0.000 description 1
- 102000004889 Interleukin-6 Human genes 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 241000581650 Ivesia Species 0.000 description 1
- 239000012097 Lipofectamine 2000 Substances 0.000 description 1
- 108091054455 MAP kinase family Proteins 0.000 description 1
- 102000043136 MAP kinase family Human genes 0.000 description 1
- 239000007993 MOPS buffer Substances 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- 102000002584 Octamer Transcription Factor-3 Human genes 0.000 description 1
- 108010068425 Octamer Transcription Factor-3 Proteins 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 108091007960 PI3Ks Proteins 0.000 description 1
- 208000031481 Pathologic Constriction Diseases 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 240000007711 Peperomia pellucida Species 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 108091008611 Protein Kinase B Proteins 0.000 description 1
- 241000382353 Pupa Species 0.000 description 1
- 238000012181 QIAquick gel extraction kit Methods 0.000 description 1
- 241000713810 Rat sarcoma virus Species 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 108020005091 Replication Origin Proteins 0.000 description 1
- 108700008625 Reporter Genes Proteins 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 102100028904 Serine/threonine-protein kinase MARK2 Human genes 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 102000009523 Transcription Factor 4 Human genes 0.000 description 1
- 108010048992 Transcription Factor 4 Proteins 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 238000010306 acid treatment Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 125000000613 asparagine group Chemical group N[C@@H](CC(N)=O)C(=O)* 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 102000005936 beta-Galactosidase Human genes 0.000 description 1
- 108010005774 beta-Galactosidase Proteins 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 210000004413 cardiac myocyte Anatomy 0.000 description 1
- 101150102092 ccdB gene Proteins 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000006059 cover glass Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000003436 cytoskeletal effect Effects 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 239000013578 denaturing buffer Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 108010030074 endodeoxyribonuclease MluI Proteins 0.000 description 1
- 238000012869 ethanol precipitation Methods 0.000 description 1
- 230000006126 farnesylation Effects 0.000 description 1
- 238000013230 female C57BL/6J mice Methods 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 238000003205 genotyping method Methods 0.000 description 1
- 230000006130 geranylgeranylation Effects 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 125000005842 heteroatom Chemical group 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229940100601 interleukin-6 Drugs 0.000 description 1
- 238000001638 lipofection Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000008531 maintenance mechanism Effects 0.000 description 1
- 238000013227 male C57BL/6J mice Methods 0.000 description 1
- 230000036244 malformation Effects 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000002205 phenol-chloroform extraction Methods 0.000 description 1
- 150000003916 phosphatidylinositol 3,4,5-trisphosphates Chemical class 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 210000001778 pluripotent stem cell Anatomy 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000008844 regulatory mechanism Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000003705 ribosome Anatomy 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 210000003765 sex chromosome Anatomy 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000010473 stable expression Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000036262 stenosis Effects 0.000 description 1
- 208000037804 stenosis Diseases 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 125000002987 valine group Chemical group [H]N([H])C([H])(C(*)=O)C([H])(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
- 239000013585 weight reducing agent Substances 0.000 description 1
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2002073138A JP4118579B2 (ja) | 2002-03-15 | 2002-03-15 | Ecat5ノックアウト細胞 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2002073138A JP4118579B2 (ja) | 2002-03-15 | 2002-03-15 | Ecat5ノックアウト細胞 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2003265166A JP2003265166A (ja) | 2003-09-24 |
| JP2003265166A5 JP2003265166A5 (enExample) | 2005-08-25 |
| JP4118579B2 true JP4118579B2 (ja) | 2008-07-16 |
Family
ID=29202943
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2002073138A Expired - Fee Related JP4118579B2 (ja) | 2002-03-15 | 2002-03-15 | Ecat5ノックアウト細胞 |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP4118579B2 (enExample) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3024338U (ja) * | 1995-11-02 | 1996-05-21 | 株式会社エレム | ア−ス端子を有するプラグ |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114845727B (zh) | 2019-12-11 | 2025-08-26 | 国立大学法人东京大学 | 基于Mycl瞬时表达的可增殖性胰岛前体细胞样细胞的诱导和胰岛素阳性细胞的分化诱导 |
| JP2024163805A (ja) | 2023-05-12 | 2024-11-22 | 国立大学法人 東京大学 | グルコース経路活性化による膵島細胞の増殖活性化 |
-
2002
- 2002-03-15 JP JP2002073138A patent/JP4118579B2/ja not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3024338U (ja) * | 1995-11-02 | 1996-05-21 | 株式会社エレム | ア−ス端子を有するプラグ |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2003265166A (ja) | 2003-09-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4901471B2 (ja) | 体細胞核初期化物質のスクリーニング方法 | |
| CA2908697C (en) | Targeted modification of rat genome | |
| WO2009063722A1 (ja) | ヒトチトクロームp450遺伝子(クラスター)を含む哺乳動物人工染色体ベクター及びそれを保持する非ヒト哺乳動物 | |
| KR20180100303A (ko) | 키메라 배아-보조 기관 생성을 위한 조성물 및 방법 | |
| JP2016129519A (ja) | 不活化された内因性遺伝子座を有するトランスジェニックニワトリ | |
| JP2007259871A (ja) | 細胞周期を仲介する組成物および方法 | |
| JP2010522565A (ja) | 胚性幹細胞分化を調整する方法 | |
| JP6204345B2 (ja) | 幹細胞の多能性を高めるための組成物及び方法 | |
| Pryzhkova et al. | Adaptation of the AID system for stem cell and transgenic mouse research | |
| CN103635588B (zh) | 用于筛选具有体重调节作用的物质的方法 | |
| JP5075641B2 (ja) | 遺伝子改変動物およびその用途 | |
| JPWO2005054463A1 (ja) | レトロトランスポゾンを用いた哺乳動物のゲノム改変技術の開発 | |
| JP7071749B2 (ja) | 分化ポテンシャルを改変した多能性幹細胞の動物の作製への応用 | |
| JP4118579B2 (ja) | Ecat5ノックアウト細胞 | |
| WO2012004346A1 (en) | Genetically engineered stem cells, method of production and uses thereof | |
| JP2011518566A (ja) | Abi1/hssh3bp1コンディショナルノックアウトマウス | |
| US7226994B2 (en) | Cell surface expressed marker of pluripotency | |
| EP1842909A1 (en) | Method for removing desired chromosome and tailor-made medical treatment utilizing the same | |
| KR20250144399A (ko) | 변형된 klhdc7b 유전자좌를 포함하는 동물 | |
| JP4510490B2 (ja) | Jab1/COP9シグナロソームを利用した細胞増殖等の制御方法 | |
| JP7203367B2 (ja) | 哺乳動物細胞用遺伝子導入ベクター | |
| KR20040062981A (ko) | 프로스타글란딘 e 합성효소 2 유전자의 붕괴 | |
| Yin | Expression and developmental requirement for transcription factor Sp2 | |
| US20030027777A1 (en) | Methods for enhancing animal growth and cell proliferation by elimination of the cyclin-dependent kinase inhibitor function of p27Kip1 | |
| JP2004344092A (ja) | Orc4遺伝子変異非ヒト動物 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20050208 |
|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20050208 |
|
| A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A712 Effective date: 20051026 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20070925 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20071126 |
|
| 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: 20080415 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20080423 |
|
| R150 | Certificate of patent or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110502 Year of fee payment: 3 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120502 Year of fee payment: 4 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120502 Year of fee payment: 4 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130502 Year of fee payment: 5 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20140502 Year of fee payment: 6 |
|
| LAPS | Cancellation because of no payment of annual fees |