DK2016194T3 - Fremgangsmåder og sammensætninger til ekspression af viralt negativ-sense-rna i hundeceller - Google Patents

Fremgangsmåder og sammensætninger til ekspression af viralt negativ-sense-rna i hundeceller Download PDF

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DK2016194T3
DK2016194T3 DK07760860.2T DK07760860T DK2016194T3 DK 2016194 T3 DK2016194 T3 DK 2016194T3 DK 07760860 T DK07760860 T DK 07760860T DK 2016194 T3 DK2016194 T3 DK 2016194T3
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virus
influenza
cells
canine
rna
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Gregory Duke
George Kemble
Zhaoti Wang
James Young
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Medimmune Llc
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Priority claimed from PCT/US2006/023867 external-priority patent/WO2007002008A2/en
Priority claimed from US11/501,067 external-priority patent/US7790434B2/en
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Claims (18)

1. Isoleret nukleinsyre, som omfatter en hunde-RNA-polymerase I-promotor, hvor nukleinsyren omfatter (a) et polynukleotid med nukleotidsekvensen ifølge SEQ ID NO: 26, (b) et funktionelt aktivt fragment af polynukleotidet som defineret i (a), hvilket fragment omfatter mindst 250 eller mindst 350 eller mindst 450 sammenhængende nukleotider fra sekvensen ifølge SEQ ID NO: 26, eller (c) et polynukleotid, som udviser mindst 99% identitet med polynukleotidet som defineret i (a).
2. Isoleret nukleinsyresekvens ifølge krav 1, som er operativt forbundet til cDNA, der koder for et viralt, genomisk negativ-streng-RNA eller det tilsvarende cRNA.
3. Isoleret nukleinsyre ifølge krav 2, hvor det virale, genomiske negativ-streng-RNA er et genomisk influenza-RNA.
4. Isoleret nukleinsyre ifølge krav 2 eller 3, hvor nukleinsyren endvidere omfatter en transskriptionstermineringssekvens.
5. Isoleret nukleinsyre ifølge krav 2, 3 eller 4, hvor cDNA'et er et influenzavirus-cDNA valgt fra gruppen bestående af: polymerase-basisprotein 2 (PB2)-cDNA, polymerase- basisprotein 1 (PBl)-cDNA, surt polymeraseprotein (PA, polymerase acidic protein)-cDNA, hæmagglutinin (HA)-cDNA, nukleoprotein (NP)-cDNA, neuraminidase (NA)-cDNA, matrixprotein 1 (Ml)-cDNA, matrixprotein 2 (M2)-cDNA, ikke- strukturprotein 1 (NSI, non-structural protein 1)- og ikke-strukturprotein 2 (NS2, non-structural protein 2)-cDNA.
6. Ekspressionsvektor, som omfatter en nukleinsyre ifølge et hvilket som helst af kravene 1-5.
7. Fremgangsmåde til frembringelse af et rekombinant influenzavirus, hvilken fremgangsmåde omfatter dyrkning af en hundecelle omfattende en ekspressionsvektor ifølge krav 6 og en eller flere ekspressionsvektorer, som udtrykker et mRNA, hvilket mRNA koder for et eller flere influenzapolypeptider valgt fra gruppen bestående af: polymerase- basisprotein 2 (PB2), polymerase-basisprotein 1 (PB1), surt polymeraseprotein (PA), hæmagglutinin (HA), nukleoprotein (NP), neuraminidase (NA), matrixprotein 1 (Ml), matrixprotein 2 (M2), ikke-strukturprotein 1 (NSI) og ikke-strukturprotein 2 (NS2), hvor mindst otte ekspressionsvektorer, som hver inkorporerer et influenzagenomsegment, hvilke influenzagenomsegmenter er forskellige, anvendes til at indføre et komplet influenzagenom i hundecellen; og isolering af det rekombinante influenzavirus.
8. Celle, som omfatter en ekspressionsvektor ifølge krav 6.
9. Fremgangsmåde til frembringelse af et genomisk influenza-RNA, hvilken fremgangsmåde omfatter indføring af ekspressionsvektoren ifølge krav 6 i hundeceller for derved at frembringe et genomisk influenza-RNA.
10. Fremgangsmåde til frembringelse af et rekombinant influenzavirus, hvilken fremgangsmåde omfatter i) indføring af ekspressionsvektorer i en population af hundeceller a) ekspression i cellerne af genomiske vRNA-segmenter for at tilvejebringe de komplette genomiske vRNA-segmenter for virusset, hvor en eller flere af ekspressionsvektorerne omfatter en nukleinsyresekvens for en hunde-RNA-polymerase I-promotor ifølge krav 1, og b) også ekspression i cellerne af mRNA, der koder for et eller flere influenzapolypeptider valgt fra gruppen bestående af: polymerase-basisprotein 2 (PB2), polymerase- basisprotein 1 (PB1), surt polymeraseprotein (PA), hæmagglutinin (HA), nukleoprotein (NP), neuraminidase (NA), matrixprotein 1 (Ml), matrixprotein 2 (M2), ikke- strukturprotein 1 (NSI) og ikke-strukturprotein 2 (NS2); og ii) dyrkning af cellerne, hvorved der frembringes influenzaviruspartikler.
11. Fremgangsmåde ifølge krav 9 eller 10, hvor hundecellerne er nyreceller.
12. Fremgangsmåde krav 10 eller 11, hvor det rekombinante influenzavirus er et reassortant virus.
13. Fremgangsmåde ifølge et hvilket som helst af kravene 10- 12, hvor frembragte influenzapartikler er infektiøse.
14. Fremgangsmåde ifølge et hvilket som helst af kravene 10- 13, hvor mRNA'et koder for hæmagglutininet (HA) og neuraminidasen (NA) fra en patogen influenzastamme.
15. Fremgangsmåde ifølge et hvilket som helst af kravene 10- 14, hvor mRNA'et koder for polymerase-basisprotein 2 (PB2), polymerase-basisprotein 1 (PB1), surt polymeraseprotein (PA), hæmagglutinin (HA), nukleoprotein (NP), neuraminidase (NA), matrixprotein 1 (Ml), matrixprotein 2 (M2), ikke- strukturprotein 1 (NSI) og ikke-strukturprotein 2 (NS2) fra en svækket influenzastamme.
16. Fremgangsmåde ifølge et hvilket som helst af kravene 10- 15, hvor det rekombinante virus er valgt fra gruppen bestående af: et kuldetilpasset influenzavirus, et svækket influenzavirus, et temperaturfølsomt influenzavirus og et svækket, temperaturfølsomt, kuldetilpasset influenzavirus.
17. Fremgangsmåde ifølge krav 16, hvor den svækkede influenzastamme er valgt fra gruppen bestående af A/Ann Arbor/6/60, B/Ann Arbor/1/66, B/Leningrad/14/17/55, B14/5/1, B/USSR/60/69, B/Leningrad/179/86, B/Leningrad/14/55, B/England/2608/76 og A/Puerto Rico/8/34.
18. Ekspressionsvektor pAD4000 ifølge SEQ ID NO: 29.
DK07760860.2T 2006-04-19 2007-04-18 Fremgangsmåder og sammensætninger til ekspression af viralt negativ-sense-rna i hundeceller DK2016194T3 (da)

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US79352206P 2006-04-19 2006-04-19
US79352506P 2006-04-19 2006-04-19
PCT/US2006/023867 WO2007002008A2 (en) 2005-06-21 2006-06-20 Methods and compositions for expressing negative-sense viral rna in canine cells
US11/455,734 US20060286591A1 (en) 2005-06-21 2006-06-20 Methods and compositions for expressing negative-sense viral RNA in canine cells
US11/501,067 US7790434B2 (en) 2005-06-21 2006-08-09 Methods and compositions for expressing negative-sense viral RNA in canine cells
PCT/US2007/066895 WO2007124327A2 (en) 2006-04-19 2007-04-18 Methods and compositions for expressing negative-sense viral rna in canine cells

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JP (1) JP5491173B2 (da)
KR (1) KR101436176B1 (da)
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AU (1) AU2007240448B8 (da)
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HK (1) HK1136849A1 (da)
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