AR123776A1 - Construcciones de ácido nucleico para la activación simultánea de genes - Google Patents

Construcciones de ácido nucleico para la activación simultánea de genes

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Publication number
AR123776A1
AR123776A1 ARP210102826A ARP210102826A AR123776A1 AR 123776 A1 AR123776 A1 AR 123776A1 AR P210102826 A ARP210102826 A AR P210102826A AR P210102826 A ARP210102826 A AR P210102826A AR 123776 A1 AR123776 A1 AR 123776A1
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Argentina
Prior art keywords
genes
dna element
recombinase
element according
dna
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ARP210102826A
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English (en)
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Simon Austaender
Ulrich Goepfert
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Hoffmann La Roche
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Publication of AR123776A1 publication Critical patent/AR123776A1/es

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    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86Viral vectors
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    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/87Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90Stable introduction of foreign DNA into chromosome
    • C12N15/902Stable introduction of foreign DNA into chromosome using homologous recombination
    • C12N15/907Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
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    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
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    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/64General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
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    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
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    • C12N2710/00011Details
    • C12N2710/10011Adenoviridae
    • C12N2710/10311Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10351Methods of production or purification of viral material
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    • C12N2750/00011Details
    • C12N2750/14011Parvoviridae
    • C12N2750/14111Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14151Methods of production or purification of viral material
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    • C12N2800/00Nucleic acids vectors
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    • C12N2830/00Vector systems having a special element relevant for transcription
    • C12N2830/50Vector systems having a special element relevant for transcription regulating RNA stability, not being an intron, e.g. poly A signal
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    • C12Q2521/00Reaction characterised by the enzymatic activity
    • C12Q2521/50Other enzymatic activities
    • C12Q2521/507Recombinase

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  • Genetics & Genomics (AREA)
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Abstract

En el presente documento se informan novedosas construcciones de ADN y procedimientos de uso de las mismas. La presente invención usa una disposición deliberada de promotores y genes no productivos / inactivos en las hebras codificantes y de molde de las moléculas de ADN, que se convierten en su forma activa mediante la interacción con una recombinasa específica de sitio. Más detalladamente, el elemento de ADN de acuerdo con la presente invención no es funcional con respecto a la expresión del primer y el segundo gen contenidos. Al no ser funcional con respecto a la expresión del primer y el segundo gen, el elemento de ADN de acuerdo con la invención se puede integrar en el genoma de una célula sin el riesgo de que los genes estructurales comprendidos se expresen ya directamente después de la integración. Los genes sólo se expresan una vez que una recombinasa que reconoce y es funcional con las secuencias de reconocimiento de recombinación del elemento de ADN se activa o se introduce en la célula. De este modo, se inicia una inversión de casete mediada por recombinasa (RMCI) entre la primera y la segunda secuencia de reconocimiento de recombinasa mutada en el elemento de ADN genómicamente integrado de la invención. La RMCI da como resultado una inversión de la parte del elemento de ADN de acuerdo con la invención que se localiza entre las dos secuencias de reconocimiento de recombinasa mutantes. De este modo, el primer promotor queda enlazado de forma funcional al primer gen y el segundo promotor queda enlazado de forma funcional al segundo gen. Sólo después de esto, el primer y el segundo gen se transcriben y se expresan las respectivas proteínas codificadas. Por tanto, el elemento de ADN de acuerdo con la presente invención es especialmente útil en la activación simultánea de dos genes dentro de una célula.
ARP210102826A 2020-10-15 2021-10-13 Construcciones de ácido nucleico para la activación simultánea de genes AR123776A1 (es)

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EP20202009 2020-10-15

Publications (1)

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AR123776A1 true AR123776A1 (es) 2023-01-11

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US (1) US20220154223A1 (es)
EP (1) EP4229204A1 (es)
JP (1) JP2023546113A (es)
KR (1) KR20230085170A (es)
CN (1) CN116348607A (es)
AR (1) AR123776A1 (es)
AU (1) AU2021363098A1 (es)
CA (1) CA3197726A1 (es)
IL (1) IL302045A (es)
MX (1) MX2023004178A (es)
TW (1) TW202229558A (es)
WO (1) WO2022079082A1 (es)

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TW202229558A (zh) 2022-08-01
CN116348607A (zh) 2023-06-27
US20220154223A1 (en) 2022-05-19
JP2023546113A (ja) 2023-11-01
MX2023004178A (es) 2023-05-03
EP4229204A1 (en) 2023-08-23
IL302045A (en) 2023-06-01
AU2021363098A1 (en) 2023-05-18
CA3197726A1 (en) 2022-04-21
WO2022079082A1 (en) 2022-04-21
KR20230085170A (ko) 2023-06-13

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