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Last Updated: October 16, 2019

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Claims for Patent: 6,579,681

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Summary for Patent: 6,579,681
Title: Test system for detecting a splicing reaction and use thereof
Abstract:The present invention relates to a test system comprising (a) one or more identical or different immobilized nucleic acid(s) having at least one spliceable nucleic acid, (b) at least one gel-free detection system for detecting a splicing reaction, where appropriate (c) at least one composition comprising splicing components, and preferably (d) suitable detection probes, and, where appropriate, (e) further aids.
Inventor(s): Huls; Christoph (Herne, DE), Bauer; Bettina (Frankfurt, DE), Simandi; Claus (Frankfurt, DE), Luhrmann; Reinhard (Marburg, DE), Achsel; Tilmann (Gottingen, DE), Vornlocher; Hans-Peter (Bayreuth, DE)
Assignee: Aventis Research and Technologies GmbH & Co. KG (Frankfurt, DE)
Application Number:09/857,063
Patent Claims:1. A test system for detecting a splicing reaction comprising (a) one or more identical or different nucleic acid(s), encompassing at least one spliceable nucleic acid, wherein the nucleic acid(s) is (are) immobilized on a solid phase, (b) at least one composition comprising splicing components, (c) at least one suitable detection probe, (d) at least one device for detecting binding of the detection probe, wherein the device does not comprise a gel.

2. The test system as claimed in claim 1 wherein the spliceable nucleic acid according to feature (a) comprises at least two exons which are separated by at least one intron.

3. The test system as claimed in claim 1 wherein the suitable detection probe according to feature (c) is a nucleic acid complementary to the spliceable nucleic acid, a low molecular weight compound which binds the spliceable nucleic acid or a peptide or protein which binds the spliceable nucleic acid.

4. The test system as claimed in claim 3 wherein the low molecular weight compound is selected from the group consisting of theophylline, xanthine and an aminoglycoside, and the nucleic acid-binding peptide or protein is an iron responsive element binding protein IBP.

5. The test system as claimed in claim 4 wherein the aminoglycoside is tobramycin.

6. The test system as claimed in claim 3 wherein the complementary nucleic acid is complementary to at least one intron, to at least one exon or to at least one exon/intron transition site.

7. The test system as claimed in claim 3 wherein the spliceable nucleic acid or the complementary nucleic acid comprises a recognition sequence for a nucleic acid-binding low molecular weight compound or for a nucleic acid-binding peptide or protein.

8. The test system as claimed in claim 7 wherein the recognition sequence for a nucleic acid-binding low molecular weight compound is an aptamer sequence.

9. The test system as claimed in claim 1 wherein the detection of the splicing reaction occurs by means of a label.

10. The test system as claimed in claim 9 wherein the label is a radio-label, a fluorescent dye, biotin, digoxigenin or an antibody.

11. The test system as claimed in claim 1 wherein the detection probe binds not directly to the spliceable nucleic acid, but to a probe-binding nucleic acid that is linked to the spliceable nucleic acid.

12. The test system as claimed in claim 11 wherein at least two different probe-binding nucleic acid sequences are linked to the spliceable nucleic acid.

13. The test system as claimed in claim 1 wherein the nucleic acid according to (a) is bound to the solid phase directly covalently or indirectly via a structural element and a binding partner of the structural element or by means of hybridization.

14. The test system as claimed in claim 13 wherein the direct covalent binding is carried out via the vicinal 2',3'-hydroxyl group of the ribose backbone of the nucleic acid.

15. The test system as claimed in claim 13 wherein the structural element is a biotin linker or a dicarboxylic acid linker.

16. The test system as claimed in claim 13 wherein the binding partner is theophylline, xanthine, an aminoglycoside or a nucleic acid-binding protein.

17. The test system as claimed in claim 16 wherein the aminoglycoside is tobramycin.

18. The test system as claimed in claim 16 wherein the nucleic acid-binding protein is IBP.

19. The test system as claimed in claim 1, wherein the solid phase is selected from the group consisting of ceramics, metal, glass and plastic.

20. The nucleic acid as claimed in claim 1, wherein at least one nucleic acid is an RNA.

21. The test system as claimed in claim 20, wherein the RNA is that can be obtained by transcription of a nucleic acid selected from the group consisting of

where (Th/To) is a sequence selected from AAGTGATACC AGCATCGTCT TGATGCCCTT GGCAGCACTT GAATT (Th; SEQ ID NO:26), or GGCTTAGTAT AGCGAGGTTT AGCTACACTC GTGCTGAGCC GAATT (To; SEQ ID NO:27).

22. The test system as claimed in claim 1 wherein the composition according to feature (b) comprise small nuclear ribonucleoprotein particle (snRNP) components and non-snRNP components.

23. The test system as claimed in claim 22 wherein the snRNP components comprise U 1, U2, U4, U5 or U6 proteins.

24. The test system as claimed in claim 1 wherein the composition according to feature (b) is a cell extract.

25. The test system as claimed in claim 24 wherein the cell extract is a eucaryotic cell extract or a nuclear extract.

26. The test system as claimed in claim 25 wherein the nuclear extract is obtained from animal cells or from fungal cells.

27. The test system as claimed in claim 26, wherein the animal cells are mammalian cells.

28. The test system as claimed in claim 26, wherein the fungal cells are yeast cells.

29. The test system as claimed in claim 1 additionally comprising a buffer solution, stabilizers or ATP.

30. A method for preparing a test system as claimed in claim 1 which comprises the steps of combining (a) one or more identical or different nucleic acid(s), encompassing at least one spliceable nucleic acid, wherein the nucleic acid(s) are immobilized on a solid phase, (b) at least one composition comprising splicing components, (c) at least one suitable detection probe, and (d) at least one device for detecting binding of the detection probe, wherein the device does not comprise a gel.

31. The test system as claimed in claim 1 wherein the device according to feature (d) is an ELISA reader or a device for detecting radioactivity.

32. An RNA that can be obtained by transcription of a nucleic acid selected from the group consisting of

Summary for Patent:   Start Trial

Foriegn Application Priority Data
Foreign Country Foreign Patent Number Foreign Patent Date
Germany199 09 156Mar 2, 1999
PCT Information
PCT FiledFebruary 25, 2000PCT Application Number:PCT/EP00/01595
PCT Publication Date:September 08, 2000PCT Publication Number:WO00/52201

Details for Patent 6,579,681

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Schering INTRON A interferon alfa-2b VIAL 103132 001 1986-06-04   Start Trial Aventis Research and Technologies GmbH & Co. KG (Frankfurt, DE) 2019-03-02 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 002 1986-06-04   Start Trial Aventis Research and Technologies GmbH & Co. KG (Frankfurt, DE) 2019-03-02 RX search
Schering INTRON A interferon alfa-2b VIAL 103132 003 1986-06-04   Start Trial Aventis Research and Technologies GmbH & Co. KG (Frankfurt, DE) 2019-03-02 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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