DK2365803T3 - Polyvalente rna-nanopartikelsammensætninger - Google Patents

Polyvalente rna-nanopartikelsammensætninger Download PDF

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DK2365803T3
DK2365803T3 DK09760436.7T DK09760436T DK2365803T3 DK 2365803 T3 DK2365803 T3 DK 2365803T3 DK 09760436 T DK09760436 T DK 09760436T DK 2365803 T3 DK2365803 T3 DK 2365803T3
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rna
nanoparticle
polynucleotide
duplex
rnase
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Chad A Mirkin
David A Giljohann
Andrew E Prigodich
Pinal C Patel
Dwight Seferos
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Univ Northwestern
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Claims (14)

1. Nanopartikelsammensætning, der omfatter: et eller flere ribonukleinsyre (RNA)-polynukleotider sammen med en nanopartikel, hvor RNA-polynukleotidet har en sekvens, der danner en dupleks under betingelser, der er passende til dannelse af dupleksen; og et yderligere polynukleotid, hvor det yderligere polynukleotid har en sekvens, der er tilstrækkeligt komplementær til en sekvens i RNA-polynukleotidet til muliggørelse af hybridisering til RNA-polynukleotidet under passende betingelser til dannelse af dupleksen; hvor dupleksen tilvejebringer et polypeptidinteraktionssted, der er placeret distalt for nanopartiklen i forhold til et midtpunkt i RNA-polynukleotidet, hvor afstanden af polypeptidinteraktionsstedet fra nanopartiklen svarer til mindst 10 nukleotider; og hvor dupleksen har mindst ét domæne i en enkeltstreng af dupleksen, der er tilstrækkeligt komplementær til en sekvens i et målpolynukleotid til muliggørelse af hybridisering af enkeltstrengen til målpolynukleotidet under passende betingelser, og hybridisering af domænet i dupleksen til sekvensen i målpolynukleotidet frembringer et substratsted, der genkendes af et polypeptid.
2. Nanopartikelsammensætning ifølge krav 1, hvor RNA- polynukleotidet er kovalent forbundet med nanopartiklen.
3. Nanopartikelsammensætning ifølge krav 1, hvor det yderligere polynukleotid er RNA.
4. Nanopartikelsammensætning ifølge krav 1, hvor det yderligere polynukleotid ikke er kovalent forbundet med nanopartiklen.
5. Nanopartikelsammensætning ifølge krav 1, hvor polypeptidinteraktionsstedet binder med et protein valgt fra gruppen, der består af RNase H, RNase D, RNase L, RNase III, Dicer, Argonaute, Argonaute2 og human immundefektvirus-transaktiverende respons-RNA-bindende protein (TRBP).
6. Nanopartikelsammensætning ifølge krav 1, hvor RNA- polynukleotidet og det yderligere polynukleotid er komplementære til hinanden over en længde, der er tilstrækkelig til muliggørelse af hybridisering.
7. Nanopartikelsammensætning ifølge krav 1, hvor RNA- polynukleotidet har en halveringstid, der er ca. 1 gang længere, ca. 2 gange længere, ca. 3 gange længere, ca. 4 gange længere, ca. 5 gange længere eller mere end en halveringstid for identisk RNA, der ikke er forbundet med en nanopartikel.
8. Nanopartikelsammensætning ifølge krav 1, der har en overfladedensitet af RNA'et på mindst ca. 2 pmol/cm2 til ca. 1000 pmol/cm2.
9. Fremgangsmåde til fremstilling af en nanopartikelsammensætning ifølge et hvilket som helst af kravene 1 til 8, der omfatter trinet lagring af en blanding af en thioleret RNA-polynukleotidduplex med nanopartiklen i en serie af opløsninger til binding af RNA-polynukleotidet til nanopartiklen, hvor hver opløsning omfatter en stigende koncentration af natriumchlorid (NaCl) i forhold til en tidligere opløsning begyndende med en første opløsning, der omfatter ca. 0,1 M NaCl.
10. Fremgangsmåde ifølge krav 9, der yderligere omfatter trinet sonikering af blandingen efter det sidste lagringstrin.
11. Fremgangsmåde ifølge krav 9, der yderligere omfatter trinet isolering af nanopartiklerne.
12. Fremgangsmåde til regulering af ekspression af et målpolynukleotid, der omfatter trinet hybridisering af målpolynukleotidet med domænet i nanopartikelsammensætningen ifølge krav 1 til frembringelse af et substratsted for et polypeptid.
13. Fremgangsmåde ifølge krav 12, hvor hybridisering resulterer i degradering af målpolynukleotidet.
14. Fremgangsmåde ifølge krav 13, hvor polypeptidet er valgt fra gruppen, der består af RNase H, RNase D, RNase L, RNase III, Dicer, Argonaute, Argonaute2 og TRBP.
DK09760436.7T 2008-11-24 2009-11-24 Polyvalente rna-nanopartikelsammensætninger DK2365803T3 (da)

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CN (2) CN106955360A (da)
AU (2) AU2009316286B2 (da)
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