DK2561078T3 - Nye strukturelt designet shrna'er - Google Patents
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- DK2561078T3 DK2561078T3 DK11772783.4T DK11772783T DK2561078T3 DK 2561078 T3 DK2561078 T3 DK 2561078T3 DK 11772783 T DK11772783 T DK 11772783T DK 2561078 T3 DK2561078 T3 DK 2561078T3
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Claims (13)
1. Short-hairpin-RNA-molekyle (shRNA) der omfatter (i) en første sekvens på 21, 22 eller 23 nucleotider som er fuldstændigt komplementær til en sekvens i et målgen, der har en anden sekvens end den-modne sekvens af miR-451 og (ii) en anden sekvens der direkte følger den første sekvens, hvilken anden sekvens er fuldstændig komplementær til sekvensen af de første 17 eller 18 nucleotider, der er talt fra 5'-enden af den første sekvens, hvor de sidste 3 eller sidste 4 nucleotider af den første sekvens danner et loopområde i shRNA'et.
2. ShRNA’et ifølge krav 1, hvor den første sekvens på 21,22 eller 23 nucleotider er komplementær til et kodende område af målgenet.
3. ShRNA ifølge krav 1 eller 2, hvor den første sekvens af 21, 22 eller 23 nucleotider er komplementær til en sekvens i et mRNA-molekyle, der er kodet af genet, hvor sekvensen i mRNA-molekylet er til stede i sekvensen af målgenet.
4. ShRNA’et ifølge krav 3, hvor den første sekvens af 21,22 eller 23 nucleotider er komplementær til en sekvens, der omfatter både et translateret område og en del af sekvensen af det 3'-utranslaterede område (UTR) i et mRNA-molekyle, der er kodet af genet, hvor 3'-UTR-sekvensen i mRNA-molekylet er til stede i sekvensen af målgenet.
5. ShRNA ifølge et hvilket som helst af kravene 1 til 4, hvor den anden sekvens der direkte følger den første sekvens, er fuldstændig komplementær til sekvensen af de første 18 nucleotider, der er talt fra 5'-enden af den første sekvens.
6. ShRNA ifølge et hvilket som helst af kravene 1 til 5, hvilket shRNA består af fra 38 til 50 nucleotider.
7. ShRNA ifølge et hvilket som helst af kravene 1 til 6 der har strukturen
hvori X2 til X22 er nucleotider, der er komplementære til en sekvens i et målgen og er i en anden sekvens end den modne sekvens af miR-451; Y4 til Y20 er nucleotider, der er komplementære til X2 til Xw; og Χι, Υι, Y2, og Y3 er nucleotider, der kan være til stede eller fraværende, hvori Xi og Y3, når de er til stede, kan være komplementære eller ikke komplementære.
8. Ekspressionsvektor der omfatter en sekvens, som koder for shRNA'et ifølge et hvilket som helst af kravene 1 til 7, der er operativt forbundet med en RNA-poly-merasepromotor.
9. Bibliotek af ekspressionsvektorer, hvor hver ekspressionsvektor koder for shRNA'et ifølge et hvilket som helst af kravene 1 til 7, der er operativt forbundet med en RNA-polymerasepromotor.
10. Isoleret pattedyrcelle der omfatter shRNA’et ifølge et hvilket som helst af kravene 1 til 7.
11. Transgent ikke-humant pattedyr, hvis genom omfatter en sekvens der koder for shRNA'et ifølge et hvilket som helst af kravene 1 til 7.
12. In v/fro-fremgangsmåde til dæmpning af ekspression af et målgen i en pattedyrcelle, hvilken fremgangsmåde omfatter at indføre en ekspressionsvektor der omfatter en sekvens, som koder for short-hairpin-RNA-molekylet (shRNA) ifølge et hvilket som helst af kravene 1 til 7, i pattedyrcellen, hvilket shRNA-molekyle er udtrykt i pattedyrcellen i en mængde, der er tilstrækkelig til at dæmpe ekspression
af målgenet på en sekvensspecifik måde, hvorved ekspression af målgenet hæmmes.
13. Ekspressionsvektoren ifølge krav 8 til anvendelse ved dæmpning af ekspression af et målgen i en pattedyrcelle.
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PCT/US2011/033615 WO2011133889A2 (en) | 2010-04-23 | 2011-04-22 | NOVEL STRUCTURALLY DESIGNED shRNAs |
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