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 proteínas

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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
Application number
BR112020000406A
Other languages
English (en)
Inventor
Aurélio Krieger Marco
Luis Kessler Rafael
Roberto Benghi Soares Luis
Preti Henrique
Su Leon
Original Assignee
Inst De Biologia Molecular Do Parana Ibmp
S P L Evolution Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Inst De Biologia Molecular Do Parana Ibmp, S P L Evolution Ltd filed Critical Inst De Biologia Molecular Do Parana Ibmp
Publication of BR112020000406A2 publication Critical patent/BR112020000406A2/pt

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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

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  • 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)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (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.
BR112020000406A 2017-07-10 2017-07-10 Plataforma genética para superexpressão heteróloga associada à seleção de células altamente produtoras de proteínas BR112020000406A2 (pt)

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)

* Cited by examiner, † Cited by third party
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
BR112022003970A2 (pt) 2019-09-03 2022-06-21 Myeloid Therapeutics Inc Métodos e composições para integração genômica

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2309000A1 (en) * 1997-11-13 1999-05-27 Regents Of The University Of Minnesota Tc1-based transposon vectors
WO2003089622A2 (en) * 2002-04-22 2003-10-30 University Of Michigan Novel genes, compositions, and methods for modulating the unfolded protein response
NZ544335A (en) * 2003-06-11 2009-01-31 Biogen Idec Inc Method to increase protein production in culture
US7514237B2 (en) * 2004-09-02 2009-04-07 Wyeth Systems and methods for protein production
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.

Also Published As

Publication number Publication date
WO2019010551A1 (pt) 2019-01-17

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Legal Events

Date Code Title Description
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]