BR112018070353A2 - linhagem de célula de mamífero para produção de proteína e geração de biblioteca - Google Patents

linhagem de célula de mamífero para produção de proteína e geração de biblioteca

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Publication number
BR112018070353A2
BR112018070353A2 BR112018070353A BR112018070353A BR112018070353A2 BR 112018070353 A2 BR112018070353 A2 BR 112018070353A2 BR 112018070353 A BR112018070353 A BR 112018070353A BR 112018070353 A BR112018070353 A BR 112018070353A BR 112018070353 A2 BR112018070353 A2 BR 112018070353A2
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Brazil
Prior art keywords
cell line
marker protein
cells
mammalian cell
protein production
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BR112018070353A
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English (en)
Inventor
PAROLA Cristina
Mason Derek
Pogson Mark
Reddy Sai
Kelton William
Original Assignee
Eth Zuerich
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Publication date
Application filed by Eth Zuerich filed Critical Eth Zuerich
Publication of BR112018070353A2 publication Critical patent/BR112018070353A2/pt

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    • 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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/102—Mutagenizing nucleic acids
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    • 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
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    • 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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
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    • 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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1058—Directional evolution of libraries, e.g. evolution of libraries is achieved by mutagenesis and screening or selection of mixed population of organisms
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • 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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1082—Preparation or screening gene libraries by chromosomal integration of polynucleotide sequences, HR-, site-specific-recombination, transposons, viral vectors
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    • 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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1086—Preparation or screening of expression libraries, e.g. reporter assays
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    • 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
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    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90—Stable introduction of foreign DNA into chromosome
    • C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
    • C12N15/907—Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0635—B lymphocytes
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    • 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
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B10/00—Directed molecular evolution of macromolecules, e.g. RNA, DNA or proteins
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    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B20/00—Methods specially adapted for identifying library members
    • C40B20/04—Identifying library members by means of a tag, label, or other readable or detectable entity associated with the library members, e.g. decoding processes
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    • C40—COMBINATORIAL TECHNOLOGY
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    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
    • C40B40/08—Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries
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    • 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]
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    • C12N2510/00—Genetically modified cells
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    • C12N2510/00—Genetically modified cells
    • C12N2510/02—Cells for production
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2800/00—Nucleic acids vectors
    • C12N2800/80—Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2521/00—Reaction characterised by the enzymatic activity
    • C12Q2521/50—Other enzymatic activities
    • C12Q2521/539—Deaminase

Abstract

de acordo com um primeiro aspecto da invenção, um método para a geração de uma linhagem celular é fornecido, compreendendo as etapas de (a) fornecer uma pluralidade de células b de mamíferos, em que cada uma dentre a pluralidade de células b compreende uma sequência de dna genômico transgênico que codifica uma proteína marcadora inserida em um locus de imunoglobulina endógena compreendido na dita célula b, e em que a sequência de dna genômico transgênico é favorável à clivagem por uma nuclease direcionada a sítio, particularmente cas9; (b) substituir a sequência de dna genômico transgênico que codifica a proteína marcadora por uma segunda sequência de dna transgênico que codifica uma proteína de interesse; (c) classificar as células b com base na presença ou ausência da proteína marcadora; e (d) coletar células b em que a proteína marcadora está ausente.
BR112018070353A 2016-04-04 2017-03-17 linhagem de célula de mamífero para produção de proteína e geração de biblioteca BR112018070353A2 (pt)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP16163734 2016-04-04
PCT/EP2017/056373 WO2017174329A1 (en) 2016-04-04 2017-03-17 Mammalian cell line for protein production and library generation

Publications (1)

Publication Number Publication Date
BR112018070353A2 true BR112018070353A2 (pt) 2019-01-29

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BR112018070353A BR112018070353A2 (pt) 2016-04-04 2017-03-17 linhagem de célula de mamífero para produção de proteína e geração de biblioteca

Country Status (9)

Country Link
US (2) US11802281B2 (pt)
EP (1) EP3440194A1 (pt)
JP (2) JP2019513415A (pt)
KR (1) KR20180132705A (pt)
CN (1) CN109072191B (pt)
BR (1) BR112018070353A2 (pt)
CA (1) CA3017678A1 (pt)
IL (2) IL299000A (pt)
WO (1) WO2017174329A1 (pt)

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