CA2573022C - Novel sequence for improving expression of nucleic acid - Google Patents
Novel sequence for improving expression of nucleic acid Download PDFInfo
- Publication number
- CA2573022C CA2573022C CA2573022A CA2573022A CA2573022C CA 2573022 C CA2573022 C CA 2573022C CA 2573022 A CA2573022 A CA 2573022A CA 2573022 A CA2573022 A CA 2573022A CA 2573022 C CA2573022 C CA 2573022C
- Authority
- CA
- Canada
- Prior art keywords
- sequence
- repressor
- nucleic acid
- sequences
- expression
- 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
- 230000014509 gene expression Effects 0.000 title claims abstract description 183
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 90
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 77
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 77
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 194
- 101710197208 Regulatory protein cro Proteins 0.000 claims abstract description 190
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 114
- 238000000034 method Methods 0.000 claims abstract description 55
- 210000004027 cell Anatomy 0.000 claims description 171
- 230000000694 effects Effects 0.000 claims description 85
- 239000012634 fragment Substances 0.000 claims description 59
- 239000003550 marker Substances 0.000 claims description 49
- 238000011144 upstream manufacturing Methods 0.000 claims description 42
- 229920001184 polypeptide Polymers 0.000 claims description 39
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 39
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 39
- 210000004978 chinese hamster ovary cell Anatomy 0.000 claims description 32
- 238000004519 manufacturing process Methods 0.000 claims description 19
- 239000002773 nucleotide Substances 0.000 claims description 14
- 125000003729 nucleotide group Chemical group 0.000 claims description 14
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 12
- 210000004962 mammalian cell Anatomy 0.000 claims description 12
- 238000012258 culturing Methods 0.000 claims description 11
- 108020004414 DNA Proteins 0.000 description 102
- 241000282414 Homo sapiens Species 0.000 description 77
- 239000013612 plasmid Substances 0.000 description 61
- 238000001890 transfection Methods 0.000 description 58
- 241000701022 Cytomegalovirus Species 0.000 description 45
- 108010084455 Zeocin Proteins 0.000 description 42
- CWCMIVBLVUHDHK-ZSNHEYEWSA-N phleomycin D1 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC[C@@H](N=1)C=1SC=C(N=1)C(=O)NCCCCNC(N)=N)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C CWCMIVBLVUHDHK-ZSNHEYEWSA-N 0.000 description 42
- 108700019146 Transgenes Proteins 0.000 description 41
- 241000829100 Macaca mulatta polyomavirus 1 Species 0.000 description 33
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 28
- 239000003623 enhancer Substances 0.000 description 23
- 102000018651 Epithelial Cell Adhesion Molecule Human genes 0.000 description 22
- 108010066687 Epithelial Cell Adhesion Molecule Proteins 0.000 description 22
- 238000013518 transcription Methods 0.000 description 18
- 230000035897 transcription Effects 0.000 description 18
- 230000014616 translation Effects 0.000 description 16
- 239000002609 medium Substances 0.000 description 15
- 239000003795 chemical substances by application Substances 0.000 description 14
- 229950010131 puromycin Drugs 0.000 description 14
- 239000013598 vector Substances 0.000 description 14
- 238000003556 assay Methods 0.000 description 11
- 239000005090 green fluorescent protein Substances 0.000 description 10
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 9
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 9
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 9
- 230000001419 dependent effect Effects 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 9
- 238000013519 translation Methods 0.000 description 9
- 108700026244 Open Reading Frames Proteins 0.000 description 8
- 230000003115 biocidal effect Effects 0.000 description 8
- 230000010354 integration Effects 0.000 description 8
- 101100118093 Drosophila melanogaster eEF1alpha2 gene Proteins 0.000 description 7
- 230000003321 amplification Effects 0.000 description 7
- 238000010367 cloning Methods 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 238000003199 nucleic acid amplification method Methods 0.000 description 7
- 230000002829 reductive effect Effects 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 239000002299 complementary DNA Substances 0.000 description 6
- 210000003527 eukaryotic cell Anatomy 0.000 description 6
- 239000012737 fresh medium Substances 0.000 description 6
- 108060003951 Immunoglobulin Proteins 0.000 description 5
- 108060001084 Luciferase Proteins 0.000 description 5
- 230000008901 benefit Effects 0.000 description 5
- 238000010276 construction Methods 0.000 description 5
- 102000037865 fusion proteins Human genes 0.000 description 5
- 108020001507 fusion proteins Proteins 0.000 description 5
- 230000030279 gene silencing Effects 0.000 description 5
- 102000018358 immunoglobulin Human genes 0.000 description 5
- 230000008488 polyadenylation Effects 0.000 description 5
- 230000002103 transcriptional effect Effects 0.000 description 5
- 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 4
- 101710196632 LexA repressor Proteins 0.000 description 4
- 239000005089 Luciferase Substances 0.000 description 4
- 239000003242 anti bacterial agent Substances 0.000 description 4
- 230000027455 binding Effects 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 239000013604 expression vector Substances 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 230000000670 limiting effect Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000002035 prolonged effect Effects 0.000 description 4
- 230000000644 propagated effect Effects 0.000 description 4
- 230000001105 regulatory effect Effects 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
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 3
- 108010034791 Heterochromatin Proteins 0.000 description 3
- 206010035226 Plasma cell myeloma Diseases 0.000 description 3
- 102100037935 Polyubiquitin-C Human genes 0.000 description 3
- 108700008625 Reporter Genes Proteins 0.000 description 3
- 108010056354 Ubiquitin C Proteins 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 210000004458 heterochromatin Anatomy 0.000 description 3
- 210000005260 human cell Anatomy 0.000 description 3
- 238000003780 insertion Methods 0.000 description 3
- 230000037431 insertion Effects 0.000 description 3
- 239000012212 insulator Substances 0.000 description 3
- 101150066555 lacZ gene Proteins 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 201000000050 myeloid neoplasm Diseases 0.000 description 3
- SGKRLCUYIXIAHR-AKNGSSGZSA-N (4s,4ar,5s,5ar,6r,12ar)-4-(dimethylamino)-1,5,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4a,5,5a,6-tetrahydro-4h-tetracene-2-carboxamide Chemical compound C1=CC=C2[C@H](C)[C@@H]([C@H](O)[C@@H]3[C@](C(O)=C(C(N)=O)C(=O)[C@H]3N(C)C)(O)C3=O)C3=C(O)C2=C1O SGKRLCUYIXIAHR-AKNGSSGZSA-N 0.000 description 2
- 108010077544 Chromatin Proteins 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 2
- 101001035782 Gallus gallus Hemoglobin subunit beta Proteins 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- 101000669028 Homo sapiens Zinc phosphodiesterase ELAC protein 2 Proteins 0.000 description 2
- 241000701024 Human betaherpesvirus 5 Species 0.000 description 2
- 108020004684 Internal Ribosome Entry Sites Proteins 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 101100311234 Oryza sativa subsp. japonica STAR1 gene Proteins 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
- 102000002508 Peptide Elongation Factors Human genes 0.000 description 2
- 108010068204 Peptide Elongation Factors Proteins 0.000 description 2
- 102000012425 Polycomb-Group Proteins Human genes 0.000 description 2
- 108020004511 Recombinant DNA Proteins 0.000 description 2
- 239000004098 Tetracycline Substances 0.000 description 2
- 108090000848 Ubiquitin Proteins 0.000 description 2
- 102000044159 Ubiquitin Human genes 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 102100039877 Zinc phosphodiesterase ELAC protein 2 Human genes 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 230000002238 attenuated effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 229930189065 blasticidin Natural products 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 210000003483 chromatin Anatomy 0.000 description 2
- 230000002759 chromosomal effect Effects 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 229960003722 doxycycline Drugs 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 238000012226 gene silencing method Methods 0.000 description 2
- 238000001415 gene therapy Methods 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 238000010348 incorporation Methods 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 229940049954 penicillin Drugs 0.000 description 2
- 230000004481 post-translational protein modification Effects 0.000 description 2
- 230000003389 potentiating effect Effects 0.000 description 2
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 238000010187 selection method Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 229960002180 tetracycline Drugs 0.000 description 2
- 229930101283 tetracycline Natural products 0.000 description 2
- 235000019364 tetracycline Nutrition 0.000 description 2
- 150000003522 tetracyclines Chemical class 0.000 description 2
- 241000701161 unidentified adenovirus Species 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 108020005544 Antisense RNA Proteins 0.000 description 1
- 241000710189 Aphthovirus Species 0.000 description 1
- 101100519158 Arabidopsis thaliana PCR2 gene Proteins 0.000 description 1
- 108010006654 Bleomycin Proteins 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 101150074155 DHFR gene Proteins 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 230000004568 DNA-binding Effects 0.000 description 1
- 108700020787 Drosophila Pc Proteins 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000710188 Encephalomyocarditis virus Species 0.000 description 1
- 102000003951 Erythropoietin Human genes 0.000 description 1
- 108090000394 Erythropoietin Proteins 0.000 description 1
- 108010023321 Factor VII Proteins 0.000 description 1
- 108010054218 Factor VIII Proteins 0.000 description 1
- 102000001690 Factor VIII Human genes 0.000 description 1
- 101100383920 Fragaria ananassa MCSI gene Proteins 0.000 description 1
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 1
- 108090000144 Human Proteins Proteins 0.000 description 1
- 102000003839 Human Proteins Human genes 0.000 description 1
- 241001135569 Human adenovirus 5 Species 0.000 description 1
- 108700002232 Immediate-Early Genes Proteins 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 description 1
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 description 1
- 102000013463 Immunoglobulin Light Chains Human genes 0.000 description 1
- 108010065825 Immunoglobulin Light Chains Proteins 0.000 description 1
- 102000012745 Immunoglobulin Subunits Human genes 0.000 description 1
- 108010079585 Immunoglobulin Subunits Proteins 0.000 description 1
- 108091036060 Linker DNA Proteins 0.000 description 1
- 239000012097 Lipofectamine 2000 Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 101100311235 Oryza sativa subsp. japonica STAR2 gene Proteins 0.000 description 1
- 101100056806 Oryza sativa subsp. japonica STAR3 gene Proteins 0.000 description 1
- 108010022429 Polycomb-Group Proteins Proteins 0.000 description 1
- 108091030071 RNAI Proteins 0.000 description 1
- 201000000582 Retinoblastoma Diseases 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 108010022394 Threonine synthase Proteins 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 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 1
- 210000004100 adrenal gland Anatomy 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 210000001776 amniocyte Anatomy 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229960001561 bleomycin Drugs 0.000 description 1
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 238000006664 bond formation reaction Methods 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000013611 chromosomal DNA Substances 0.000 description 1
- 239000013599 cloning vector Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000003184 complementary RNA Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 102000004419 dihydrofolate reductase Human genes 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000003292 diminished effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000000267 erythroid cell Anatomy 0.000 description 1
- 229940105423 erythropoietin Drugs 0.000 description 1
- 229960000301 factor viii Drugs 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012909 foetal bovine serum Substances 0.000 description 1
- 238000002825 functional assay Methods 0.000 description 1
- 238000011990 functional testing Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 230000009368 gene silencing by RNA Effects 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 230000009422 growth inhibiting effect Effects 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 201000008968 osteosarcoma Diseases 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 230000029279 positive regulation of transcription, DNA-dependent Effects 0.000 description 1
- OXCMYAYHXIHQOA-UHFFFAOYSA-N potassium;[2-butyl-5-chloro-3-[[4-[2-(1,2,4-triaza-3-azanidacyclopenta-1,4-dien-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound [K+].CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=N[N-]N=N2)C=C1 OXCMYAYHXIHQOA-UHFFFAOYSA-N 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 235000008160 pyridoxine Nutrition 0.000 description 1
- 239000011677 pyridoxine Substances 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 238000009877 rendering Methods 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 230000001718 repressive effect Effects 0.000 description 1
- 210000001525 retina Anatomy 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 230000037426 transcriptional repression Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000030787 translation reinitiation Effects 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4702—Regulators; Modulating 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
- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/67—General methods for enhancing the 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
- Peptides Or Proteins (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US58700704P | 2004-07-08 | 2004-07-08 | |
| US60/587,007 | 2004-07-08 | ||
| EPPCT/EP2004/051405 | 2004-07-08 | ||
| EP2004051405 | 2004-07-08 | ||
| PCT/EP2005/053251 WO2006005718A2 (en) | 2004-07-08 | 2005-07-07 | Novel sequence for improving expression of nucleic acid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2573022A1 CA2573022A1 (en) | 2006-01-19 |
| CA2573022C true CA2573022C (en) | 2012-03-06 |
Family
ID=34971366
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2573022A Expired - Fee Related CA2573022C (en) | 2004-07-08 | 2005-07-07 | Novel sequence for improving expression of nucleic acid |
Country Status (19)
| Country | Link |
|---|---|
| US (1) | US7825232B2 (enExample) |
| JP (1) | JP4805262B2 (enExample) |
| KR (1) | KR101188013B1 (enExample) |
| CN (1) | CN1981057B (enExample) |
| AT (1) | ATE378426T1 (enExample) |
| AU (1) | AU2005261699B2 (enExample) |
| BR (1) | BRPI0512798A (enExample) |
| CA (1) | CA2573022C (enExample) |
| DE (1) | DE602005003369T2 (enExample) |
| DK (1) | DK1763586T3 (enExample) |
| EA (1) | EA011747B1 (enExample) |
| ES (1) | ES2296201T3 (enExample) |
| IL (1) | IL180531A (enExample) |
| MX (1) | MXPA06014553A (enExample) |
| NO (1) | NO341862B1 (enExample) |
| NZ (1) | NZ551510A (enExample) |
| PT (1) | PT1763586E (enExample) |
| WO (1) | WO2006005718A2 (enExample) |
| ZA (1) | ZA200610470B (enExample) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004056986A2 (en) * | 2002-12-20 | 2004-07-08 | Chromagenics B.V. | Means and methods for producing a protein through chromatin openers that are capable of rendering chromatin more accessible to transcription factors |
| US20100136616A1 (en) * | 2004-11-08 | 2010-06-03 | Chromagenics B.V. | Selection of Host Cells Expressing Protein at High Levels |
| JP5291341B2 (ja) * | 2004-11-08 | 2013-09-18 | クロマジェニックス ベー ヴェー | タンパク質を高レベルで発現する宿主細胞の選定 |
| US8999667B2 (en) * | 2004-11-08 | 2015-04-07 | Chromagenics B.V. | Selection of host cells expressing protein at high levels |
| US20060195935A1 (en) | 2004-11-08 | 2006-08-31 | Chromagenics B.V. | Selection of host cells expressing protein at high levels |
| US8039230B2 (en) | 2004-11-08 | 2011-10-18 | Chromagenics B.V. | Selection of host cells expressing protein at high levels |
| SI1809750T1 (sl) * | 2004-11-08 | 2012-08-31 | Chromagenics Bv | Izbira gostiteljskih celic, ki imajo visok nivo izraĹľanja proteina |
| BRPI1013141A2 (pt) | 2009-06-15 | 2015-10-06 | Cellagenics B V | novos marcadores selecionaveis rigorosos |
| CN103038352B (zh) | 2010-06-15 | 2015-12-16 | 萨拉基尼克有限公司 | 用于增强基因表达的新型基因间元件 |
| WO2012030218A1 (en) | 2010-09-01 | 2012-03-08 | Cellagenics B.V. | Nucleic acid fragments from a ribosomal protein promoter for enhancing gene expression |
| CN110484563B (zh) * | 2019-07-25 | 2023-04-07 | 新乡医学院 | 哺乳动物细胞组合表达载体、表达系统、制备方法和应用 |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ATE466941T1 (de) | 2001-07-04 | 2010-05-15 | Chromagenics Bv | Dns-sequenzen mit anti-repressor-aktivität |
| NZ537073A (en) | 2002-06-14 | 2006-08-31 | Chromagenics B | A method for simultaneous production of multiple proteins; vectors and cells for use therein |
| WO2004056986A2 (en) | 2002-12-20 | 2004-07-08 | Chromagenics B.V. | Means and methods for producing a protein through chromatin openers that are capable of rendering chromatin more accessible to transcription factors |
-
2005
- 2005-07-07 BR BRPI0512798-0A patent/BRPI0512798A/pt not_active Application Discontinuation
- 2005-07-07 EA EA200700233A patent/EA011747B1/ru not_active IP Right Cessation
- 2005-07-07 KR KR1020067027165A patent/KR101188013B1/ko not_active Expired - Fee Related
- 2005-07-07 ES ES05756870T patent/ES2296201T3/es not_active Expired - Lifetime
- 2005-07-07 US US11/632,012 patent/US7825232B2/en not_active Expired - Fee Related
- 2005-07-07 CN CN2005800229700A patent/CN1981057B/zh not_active Expired - Fee Related
- 2005-07-07 JP JP2007519805A patent/JP4805262B2/ja not_active Expired - Fee Related
- 2005-07-07 WO PCT/EP2005/053251 patent/WO2006005718A2/en not_active Ceased
- 2005-07-07 AT AT05756870T patent/ATE378426T1/de active
- 2005-07-07 CA CA2573022A patent/CA2573022C/en not_active Expired - Fee Related
- 2005-07-07 PT PT05756870T patent/PT1763586E/pt unknown
- 2005-07-07 AU AU2005261699A patent/AU2005261699B2/en not_active Ceased
- 2005-07-07 DE DE602005003369T patent/DE602005003369T2/de not_active Expired - Lifetime
- 2005-07-07 NZ NZ551510A patent/NZ551510A/en not_active IP Right Cessation
- 2005-07-07 DK DK05756870T patent/DK1763586T3/da active
- 2005-07-07 MX MXPA06014553A patent/MXPA06014553A/es active IP Right Grant
-
2006
- 2006-12-13 ZA ZA200610470A patent/ZA200610470B/xx unknown
-
2007
- 2007-01-03 IL IL180531A patent/IL180531A/en active IP Right Grant
- 2007-02-08 NO NO20070757A patent/NO341862B1/no not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| WO2006005718A3 (en) | 2006-08-17 |
| KR101188013B1 (ko) | 2012-10-08 |
| CA2573022A1 (en) | 2006-01-19 |
| HK1097004A1 (en) | 2007-06-15 |
| KR20070042122A (ko) | 2007-04-20 |
| WO2006005718A2 (en) | 2006-01-19 |
| DK1763586T3 (da) | 2008-03-17 |
| IL180531A (en) | 2011-04-28 |
| MXPA06014553A (es) | 2007-03-12 |
| US20070212755A1 (en) | 2007-09-13 |
| CN1981057B (zh) | 2012-07-04 |
| EA011747B1 (ru) | 2009-06-30 |
| PT1763586E (pt) | 2008-02-25 |
| AU2005261699B2 (en) | 2009-07-23 |
| BRPI0512798A (pt) | 2008-04-08 |
| ZA200610470B (en) | 2008-09-25 |
| CN1981057A (zh) | 2007-06-13 |
| DE602005003369D1 (de) | 2007-12-27 |
| JP2008505626A (ja) | 2008-02-28 |
| NO20070757L (no) | 2007-02-08 |
| ATE378426T1 (de) | 2007-11-15 |
| NO341862B1 (no) | 2018-02-12 |
| ES2296201T3 (es) | 2008-04-16 |
| DE602005003369T2 (de) | 2009-03-05 |
| US7825232B2 (en) | 2010-11-02 |
| IL180531A0 (en) | 2007-06-03 |
| NZ551510A (en) | 2009-10-30 |
| AU2005261699A1 (en) | 2006-01-19 |
| EA200700233A1 (ru) | 2007-06-29 |
| JP4805262B2 (ja) | 2011-11-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2581422C (en) | Selection of host cells expressing protein at high levels | |
| KR101328300B1 (ko) | 고수준으로 단백질을 발현하는 숙주세포의 선택 | |
| NZ553700A (en) | Selection of host cells expressing protein at high levels using a selectable marker with a non-optimal start codon | |
| KR100942117B1 (ko) | 유전자 전사 조절 성질을 포함하는 디엔에이 서열 및 이러한 디엔에이 서열을 탐지하고 이용하는 방법 | |
| CA2573022C (en) | Novel sequence for improving expression of nucleic acid | |
| DK2443239T3 (en) | New stringent selection markers | |
| Boccuni et al. | The human L (3) MBT polycomb group protein is a transcriptional repressor and interacts physically and functionally with TEL (ETV6) | |
| EP2611915A1 (en) | Nucleic acid fragments from a ribosomal protein promoter for enhancing gene expression | |
| KR100951759B1 (ko) | 유전자 전사 조절 성질을 포함하는 디엔에이 서열 및이러한 디엔에이 서열을 탐지하고 이용하는 방법 | |
| HK1128490B (en) | Selection of host cells expressing protein at high levels | |
| US6635751B1 (en) | Isolated nucleic acids encoding activated and suppressive forms of ATF6 | |
| US6346606B1 (en) | Protein containing a scavenger receptor cysteine rich domain | |
| US6355775B1 (en) | Transcriptional inhibitor | |
| EP1763586B1 (en) | Novel sequence for improving expression of nucleic acid | |
| HK1097004B (en) | Novel sequence for improving expression of nucleic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |
Effective date: 20220707 |