DK2327786T3 - Stedspecifikke serinrekombinaser og fremgangsmåder til anvendelse deraf - Google Patents

Stedspecifikke serinrekombinaser og fremgangsmåder til anvendelse deraf Download PDF

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DK2327786T3
DK2327786T3 DK10181444.0T DK10181444T DK2327786T3 DK 2327786 T3 DK2327786 T3 DK 2327786T3 DK 10181444 T DK10181444 T DK 10181444T DK 2327786 T3 DK2327786 T3 DK 2327786T3
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recombination
recombinase
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Malla Padidam
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Intrexon Corp
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Claims (10)

1. Fremgangsmåde til opnåelse af stedspecifik rekombination i genomisk DNA i en isoleret mammaliacelle, hvilken fremgangsmåde omfatter: tilvejebringelse af en mammaliacelle, der omfatter et første rekombinationssted og et andet rekombinationssted; etablering af kontakt mellem det første og andet rekombinationssted og et Bacillus subtilis SpPc2-fag-rekombinasepolypeptid, der kodes af et polynukleotid, der omfatter en nukleinsyre, der er mindst 90 % identisk med nukleinsyresekvensen ifølge SEQ ID NO: 1, til opnåelse af rekombination mellem rekombinationsstederne, hvor rekombinasepolypeptidet kan mediere rekombination mellem det første og andet rekombinationssted, hvor det første rekombinationssted er et fag-genomisk rekombinationsbindingssted (attP) eller et bakterie-genomisk rekombinationsbindingssted (attB), og det andet rekombinationssted er attB eller att P i genomet i den isolerede mammaliacelle, forudsat at når det første rekombinationsbindingssted er attB, er det andet rekombinationsbindingssted attP, og når det første rekombinationsbindingssted er attP, er det andet rekombinationsbindingssted attB.
2. Fremgangsmåde ifølge krav 1, hvor polynukleotidet er operabelt koblet til en promotor, der medierer ekspression af rekombinasen i mammaliacellen.
3. Fremgangsmåde ifølge krav 1, hvor rekombinasepolypeptidet indføres i mammaliacellen ved ekspression af et polynukleotid, der koder for rekombinasepolypeptidet.
4. Fremgangsmåde ifølge krav 1, hvor rekombinasepolypeptidet indføres i mammaliacellen som et polypeptid.
5. Fremgangsmåde ifølge krav 1, hvor rekombinasepolypeptidet indføres i mammaliacellen ved hjælp af messenger-RNA, der koder for rekombinasepolypeptidet.
6. Fremgangsmåde ifølge krav 1, hvor den stedspecifikke rekombination resulterer i integration, deletion, inversion, translokation eller udveksling af DNA.
7. Fremgangsmåde ifølge krav 1, hvor et polynukleotid som et resultat af rekombinationen integreres stabilt i den isolerede mammaliacelle.
8. Fremgangsmåde ifølge krav 1, der yderligere omfatter tilvejebringelse af et tredje rekombinationssted og et fjerde rekombinationssted til mammaliacellen; etablering af kontakt mellem det tredje og fjerde rekombinationssted og et andet prokaryot rekombinasepolypeptid til opnåelse af rekombination mellem det tredje og fjerde rekombinationssted, hvor det andet rekombinasepolypeptid kan mediere rekombination mellem det tredje og fjerde rekombinationssted.
9. Fremgangsmåde ifølge krav 1, hvilken fremgangsmåde anvender en vektor, der omfatter et polynukleotid af interesse, og et andet rekombinations-attB- eller attP-sted, hvor det andet rekombinations-attB- eller attP-sted omfatter en polynukleotidsekvens, der rekombinerer med et første rekombinations-attP- eller attB-sted i genomet i den isolerede mammaliacelle, og rekombinationen finder sted i nærvær af en Ξρβο2-ίagrekombinase.
10. Fremgangsmåde ifølge krav 1, hvor rekombinasepolypeptidet kodes af SEQ ID NO: 1.
DK10181444.0T 2005-02-02 2005-02-08 Stedspecifikke serinrekombinaser og fremgangsmåder til anvendelse deraf DK2327786T3 (da)

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US11/049,552 US9034650B2 (en) 2005-02-02 2005-02-02 Site-specific serine recombinases and methods of their use
EP05722807.4A EP1844155B1 (en) 2005-02-02 2005-02-08 Site-specific serine recombinases and methods of their use

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US (5) US9034650B2 (da)
EP (3) EP2327786B1 (da)
JP (4) JP5684445B2 (da)
KR (1) KR20080003780A (da)
CN (2) CN102994492B (da)
AU (1) AU2005326917B2 (da)
BR (1) BRPI0519865A2 (da)
CA (4) CA3048118A1 (da)
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ES (2) ES2584911T3 (da)
HK (1) HK1107119A1 (da)
IL (3) IL185027A (da)
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