BR112020000406A2 - Plataforma genética para superexpressão heteróloga associada à seleção de células altamente produtoras de proteínas - Google Patents
Plataforma genética para superexpressão heteróloga associada à seleção de células altamente produtoras de proteínasInfo
- Publication number
- BR112020000406A2 BR112020000406A2 BR112020000406A BR112020000406A BR112020000406A2 BR 112020000406 A2 BR112020000406 A2 BR 112020000406A2 BR 112020000406 A BR112020000406 A BR 112020000406A BR 112020000406 A BR112020000406 A BR 112020000406A BR 112020000406 A2 BR112020000406 A2 BR 112020000406A2
- Authority
- BR
- Brazil
- Prior art keywords
- selection
- proteins
- atf6
- xbp1
- producing cells
- Prior art date
Links
- 108090000623 proteins and genes Proteins 0.000 title abstract 6
- 102000004169 proteins and genes Human genes 0.000 title abstract 5
- 230000002018 overexpression Effects 0.000 title abstract 4
- 230000002068 genetic effect Effects 0.000 title abstract 2
- 210000004027 cell Anatomy 0.000 abstract 3
- 239000004098 Tetracycline Substances 0.000 abstract 2
- 238000010276 construction Methods 0.000 abstract 2
- 210000002472 endoplasmic reticulum Anatomy 0.000 abstract 2
- 230000014509 gene expression Effects 0.000 abstract 2
- 229960002180 tetracycline Drugs 0.000 abstract 2
- 229930101283 tetracycline Natural products 0.000 abstract 2
- 235000019364 tetracycline Nutrition 0.000 abstract 2
- 150000003522 tetracyclines Chemical class 0.000 abstract 2
- 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 abstract 1
- 101150014641 Atf6 gene Proteins 0.000 abstract 1
- 241000699802 Cricetulus griseus Species 0.000 abstract 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 abstract 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 abstract 1
- 102000008165 X-Box Binding Protein 1 Human genes 0.000 abstract 1
- 108010035430 X-Box Binding Protein 1 Proteins 0.000 abstract 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 abstract 1
- 230000004186 co-expression Effects 0.000 abstract 1
- 229960003722 doxycycline Drugs 0.000 abstract 1
- 239000007850 fluorescent dye Substances 0.000 abstract 1
- 230000004927 fusion Effects 0.000 abstract 1
- 230000010354 integration Effects 0.000 abstract 1
- 238000002372 labelling Methods 0.000 abstract 1
- 238000012544 monitoring process Methods 0.000 abstract 1
- 229920001184 polypeptide Polymers 0.000 abstract 1
- 102000004196 processed proteins & peptides Human genes 0.000 abstract 1
- 108090000765 processed proteins & peptides Proteins 0.000 abstract 1
- 230000001105 regulatory effect Effects 0.000 abstract 1
- 230000003248 secreting effect Effects 0.000 abstract 1
- 239000013598 vector Substances 0.000 abstract 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
-
- 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
- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Cell Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
PLATAFORMA GENÉTICA PARA SUPEREXPRESSÃO HETERÓLOGA ASSOCIADA À SELEÇÃO DE CÉLULAS ALTAMENTE PRODUTORAS DE PROTEÍNAS. A presente invenção descreve i) a construção de vetores de integração para superexpressão heteróloga associados a um sistema de seleção de células e monitoramento em tempo real da produtividade pela co-expressão de proteínas repórteres, ii) a seleção de células altamente secretoras da proteína de interesse pela marcação com sondas fluorescentes específicas para o retículo endoplasmático (RE), iii) construção de uma linhagem geneticamente modificada de células CHO para expressão condicional de proteínas de interesse sobre o controle de tetraciclina/doxiciclina (sistema TetON), iv) desenvolvimento de um gene sintético contendo as formas ativas das proteínas Atf6 e Xbp1 fusionadas em um único polipeptídeo e de linhagens de células CHO expressando a fusão Atf6-Xbp1 sob o controle (ou não) de tetraciclina, v) super expressão e alta produtividade de proteínas recombinantes pela co-expressão regulada de Atf6-Xbp1.
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/BR2017/050184 WO2019010551A1 (pt) | 2017-07-10 | 2017-07-10 | Plataforma genética para superexpressão heteróloga associada à seleção de células altamente produtoras de proteínas |
Publications (1)
Publication Number | Publication Date |
---|---|
BR112020000406A2 true BR112020000406A2 (pt) | 2022-09-13 |
Family
ID=65001063
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
BR112020000406A BR112020000406A2 (pt) | 2017-07-10 | 2017-07-10 | Plataforma genética para superexpressão heteróloga associada à seleção de células altamente produtoras de proteínas |
Country Status (2)
Country | Link |
---|---|
BR (1) | BR112020000406A2 (pt) |
WO (1) | WO2019010551A1 (pt) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108728477B (zh) * | 2017-04-24 | 2022-02-22 | 华东理工大学 | 一种高效的转座突变系统及构建方法 |
KR20220097875A (ko) | 2019-09-03 | 2022-07-08 | 마이얼로이드 테라퓨틱스, 인크. | 게놈 통합을 위한 방법 및 조성물 |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU1410399A (en) * | 1997-11-13 | 1999-06-07 | Regents Of The University Of Minnesota | Nucleic acid transfer vector for the introduction of nucleic acid into the dna of a cell |
AU2003234198A1 (en) * | 2002-04-22 | 2003-11-03 | University Of Michigan | Novel genes, compositions, and methods for modulating the unfolded protein response |
EP1639095A2 (en) * | 2003-06-11 | 2006-03-29 | Biogen Idec MA Inc. | Method to increase protein production in culture |
CA2577073C (en) * | 2004-09-02 | 2016-10-11 | Wyeth | Systems and methods for protein production employing polypeptides functional in unfolded protein response pathway |
WO2010099296A1 (en) * | 2009-02-26 | 2010-09-02 | Transposagen Biopharmaceuticals, Inc. | Hyperactive piggybac transposases |
CL2015003242A1 (es) * | 2015-11-04 | 2016-10-14 | Univ Chile | Virus aav/upr-plus, proteína de fusión upr-plus, método de tratamiento genético y su uso en el tratamiento enfermedades neurodegenerativas, tales como las enfermedades de parkinson y huntington, entre otras. |
-
2017
- 2017-07-10 WO PCT/BR2017/050184 patent/WO2019010551A1/pt active Application Filing
- 2017-07-10 BR BR112020000406A patent/BR112020000406A2/pt unknown
Also Published As
Publication number | Publication date |
---|---|
WO2019010551A1 (pt) | 2019-01-17 |
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B12F | Other appeals [chapter 12.6 patent gazette] | ||
B06W | Patent application suspended after preliminary examination (for patents with searches from other patent authorities) chapter 6.23 patent gazette] | ||
B07A | Application suspended after technical examination (opinion) [chapter 7.1 patent gazette] |