Title:
GENE EXPRESSION IN EUKARYOTIC CELLS
Document Type and Number:
WIPO Patent Application WO/2000/063359
Kind Code:
A2
Abstract:
A polynucleotide comprising a coding sequence, which coding sequence comprises a heterologous intron, for use in a method of treatment of the human or animal body by therapy. A cell, animal or plant comprising the polynucleotide and methods of making such cells, animals or plants are also provided.
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Inventors:
ROES JURGEN THEODOR (GB)
Application Number:
PCT/GB2000/001454
Publication Date:
October 26, 2000
Filing Date:
April 17, 2000
Export Citation:
Assignee:
UNIV LONDON (GB)
ROES JURGEN THEODOR (GB)
ROES JURGEN THEODOR (GB)
International Classes:
C12N9/00; (IPC1-7): C12N15/10; A01H1/00; A01K67/027; A61K48/00; C07K14/47; C12N5/06; C12N5/10; C12N15/13; C12N15/63; C12N15/67; C12N15/82
Domestic Patent References:
| WO1998040519A1 | 1998-09-17 | |||
| WO1998048004A1 | 1998-10-29 | |||
| WO1998056921A1 | 1998-12-17 | |||
| WO1995030000A1 | 1995-11-09 | |||
| WO2000000629A1 | 2000-01-06 | |||
| WO2000011200A2 | 2000-03-02 |
Foreign References:
| EP0459643A2 | 1991-12-04 |
Other References:
HASELOFF J ET AL: "REMOVAL OF A CRYPTIC INTRON AND SUBCELLULAR LOCALIZATION OF GREEN FLUORESCENT PROTEIN ARE REQUIRED TO MARK TRANSGENIC ARABIDOPSIS PLANTS BRIGHTLY" PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA,US,NATIONAL ACADEMY OF SCIENCE. WASHINGTON, vol. 94, 1 March 1997 (1997-03-01), pages 2122-2127, XP002043800 ISSN: 0027-8424
Attorney, Agent or Firm:
Woods, Geoffrey Corlett (J.A. Kemp & Co. 14 South Square Gray's Inn
London WC1R 5LX, GB)
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Claims:
CLAIMS
| 1. | A polynucleotide comprising a coding sequence, which coding sequence comprises a heterologous intron, for use in a method of treatment of the human or animal body by therapy. |
| 2. | A polynucleotide according to claim 1 for use in a method wherein the polynucleotide is stably introduced into cells of the human or animal. |
| 3. | A polynucleotide according to claim 1 or 2 wherein the coding sequence does not naturally have introns. |
| 4. | A polynucleotide according to claim 1 or 2 wherein the coding sequence comprises a cDNA sequence corresponding to a genomic DNA that comprises introns. |
| 5. | A polynucleotide according to any one of the preceding claims wherein the heterologous intron is present in a cryptic splice site. |
| 6. | A polynucleotide according to any one of the preceding claims wherein (a) a cryptic splice site which occurs in the natural form of the coding sequence has been mutated to weaken or inactivate the splice site, and/or (b) comprises a mutated splice site which has a greater splice activity than the naturally occurring form of the splice site, and/or (c) comprises a minimised form of a homologous intron. |
| 7. | A polynucleotide according to any one of the preceding claims wherein the heterologous intron is an immunoglobulin gene intron. |
| 8. | A polynucleotide according to any one of the preceding claims wherein the coding sequence encodes a therapeutic product. |
| 9. | A polynucleotide according to any one of the preceding claims which further comprises control sequences operably linked to the coding sequence, said control sequences being capable of expressing the coding sequence in the cells of the human or animal. |
| 10. | Use of a polynucleotide as defined in any one of the preceding claims in the manufacture of a medicament for treating a disease in a human or animal by stable introduction of the polynucleotide into the cells of the human or animal. |
| 11. | A polynucleotide as defined in any one of claims 1 to 9. |
| 12. | A polynucleotide according to claim 11, which is a polynucleotide as defined in any one of claims 5 to 8. |
| 13. | A polynucleotide according to claim 11 or 12 wherein the coding sequence encodes a functional Cre recombinase or GFP protein. |
| 14. | A vector comprising a polynucleotide as defined in claim 11,12 or 13. |
| 15. | A vector according to claim 14 which is a gene therapy vector. |
| 16. | A eukaryotic cell that comprises a polynucleotide or vector as defined in any one of claims 11 to 15. |
| 17. | A eukaryotic cell according to claim 16 wherein the polynucleotide or vector is integrated into the genome of the cell. |
| 18. | A cell according to claim 16 or 17 for use in a method of treatment of the human or animal body by therapy. |
| 19. | Method of producing a eukaryotic cell which is capable of expressing a gene product comprising: (i) introducing into the cell a polynucleotide or vector according to any one of claims 11 to 15, the coding sequence of which polynucleotide encodes said gene product, and optionally (ii) allowing the polynucleotide to integrate into the genome of the cell. |
| 20. | Method of producing a product comprising culturing a cell according to claim 16 or 17 in which the polynucleotide encodes the product, under conditions which allow expression of the product, and optionally recovering the product from the cell. |
| 21. | Method according to claim 20 wherein the product is a polypeptide. |
| 22. | A product made by the method of claim 20 or 21. |
| 23. | Method of producing an animal that expresses a product comprising introducing a polynucleotide or vector according to any one of claims 11 to 15 whose coding sequence encodes the product into an embryo stem cell and allowing the embryo stem cell to grow into an animal. |
| 24. | An animal or animal part, plant or plant part, plant seed or callus which comprises (i) polynucleotide or vector according to any one of claims 11 to 15 integrated into its genome, or (ii) a cell as defined in claim 16 or 17. |
| 25. | A method of obtaining a transgenic plant cell comprising: (a) transforming a plant cell with a polynucleotide or vector as defined in any one of claims 11 to 14. |
| 26. | A method of obtaining a firstgeneration transgenic plant comprising: (b) regenerating a transgenic plant cell transformed with a polynucleotide or vector as defined in any one of claims 11 to 14 to give a transgenic plant. |
| 27. | A method of obtaining a transgenic plant seed comprising: (c) obtaining a transgenic seed from a transgenic plant obtainable by step (b) of claim 26. |
| 28. | A method of obtaining a transgenic progeny plant comprising obtaining a second generation transgenic progeny plant from a firstgeneration transgenic plant obtainable by a method according to claim 27, and optionally obtaining transgenic plants of one or more further generations from the secondgeneration progeny plant thus obtained. |
| 29. | A method according to claim 28 comprising : (d) obtaining a transgenic seed from a firstgeneration transgenic plant obtainable by the method according to claim 26, then obtaining a second generation transgenic progeny plant from the transgenic seed; and/or (e) propagating clonally a firstgeneration transgenic plant obtainable by the method according to claim 26 to give a secondgeneration progeny plant; and/or (f) crossing a firstgeneration transgenic plant obtainable by a method according to claim 26 with another plant to give a secondgeneration progeny plant; and optionally (g) obtaining transgenic progeny plants of one or more further generations from the secondgeneration progeny plants thus obtained. |
| 30. | A transgenic plant cell, plant, plant seed or progeny plant obtainable by a method according to any one of claims 25 to 29. |
| 31. | A callus according to claim 24 obtainable from a transgenic plant cell, first generation plant, plant seed or progeny plant produced in a method according to any one of claims 25 to 29. |
| 32. | Use of a polynucleotide as defined in or vector as defined in any one of claims 11 to 14 to generate a transgenic plant having increased resistance to an external stress, by comparison with an equivalent nontransgenic plant. |
| 33. | Use according to claim 32 wherein the external stress is selected from a herbicide, a pathogen or pest, or an unfavourable environmental factor. |
| 34. | Use of a heterologous intron to increase the expression of a coding sequence. |
| 35. | Use according to claim 34 wherein the intron and coding sequence are present on a polynucleotide or vector as defined in any one claims 11 to 15. |
| 36. | Method of treating or preventing a disease in a human or animal comprising administering to the human or animal a polynucleotide, vector or cell as defined in any one of claims 11 to 17. |
Description:
INTERNATIONAL SEARCH REPORT Intern, al Application No PCT/GB 00/01454 C. (Continuation) DOCUMENTS CONSIDERED TO BE RELEVANT Category Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. A EP 0 459 643 A (COMMW SCIENT IND RES ORG 1,3,14, ; LUBRIZOL GENETICS INC (US)) 24-34 4 December 1991 (1991-12-04) page 4, line 33-page 5, line 40; examples 1-4 A WO 98 56921 A (DOW AGROSCIENCES LLC) 1,3,14, 17 December 1998 (1998-12-17) 24-34 page 5, line 6-13 page 7, line 1-page 8, line 14 page 12, line 28-page 31, line 29; examples6-12,17-22 page 71, line 17-page 74, line 15; examples27,28 A WO 95 30000 A (BIOTECH & BIOLOG SCIEN RES 1,6,8 ; CLARK ANTHONY JOHN (G8)) 9 November 1995 (1995-11-09) the whole document A HASELOFF J ET AL :"REMOVAL OF A CRYPTIC 1,6 INTRON AND SUBCELLULAR LOCALIZATION OF GREEN FLUORESCENT PROTEIN ARE REQUIRED TO MARK TRANSGENIC ARABIDOPSIS PLANTS BRIGHTLY" PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA, US, NATIONAL ACADEMY OF SCIENCE. WASHINGTON, vol. 94,1 March 1997 (1997-03-01), pages 2122-2127, XP002043800 ISSN: 0027-8424 the whole document P, X WO 00 00629 A (KIM JONG MOOK; YU SEUNG 1,2,8-22 SHIN (KR); KIM SUNYOUNG (KR); VIROMED LIMI) 6 January 2000 (2000-01-06) abstract page 5, line 15-page 6, line 20 page 13, line 7-page 16, line 8; example 3 P, X WO 00 11200 A (MONSANTO CO) 1,3,14, 2 March 2000 (2000-03-02) 24-34 abstract examples 1-3 1 INTERNATIONAL SEARCH REPORT Intern., al Application No . rmation on patent family members PCT/GB 00/01454 Patent document Publication Patent family Publication cited in search report date member (s) date WO 9840519 A 17-09-1998 AU 6700598 A 29-09-1998 WO 9848004 A 29-10-1998 AU 7359298 A 13-11-1998 EP 0459643 A 04-12-1991 AT 195554 T 15-09-2000 AU 643521 B 18-11-1993 AU 7710791 A 21-11-1991 CA 2042831 A 19-11-1991 CN 1063506 A 12-08-1992 DE 69132366 D 21-09-2000 JP 7067645 A 14-03-1995 US 5290924 A 01-03-1994 WO 9856921 A 17-12-1998 AU 7830198 A 30-12-1998 EP 0991764 A 12-04-2000 WO 9530000 A 09-11-1995 AU 686375 B 05-02-1998 AU 2317095 A 29-11-1995 CA 2189438 A 09-11-1995 CN 1149317 A 07-05-1997 EP 0763108 A 19-03-1997 FI 964423 A 04-11-1996 JP 9512430 T 16-12-1997 NO 964628 A 03-01-1997 NZ 284550 A 27-05-1998 US 6046380 A 04-04-2000 WO 0000629 A 06-01-2000 AU 4655499 A 17-01-2000 EP 1032697 A 06-09-2000 WO 0011200 A 02-03-2000 AU 5571699 A 14-03-2000 AU 5571999 A 14-03-2000 AU 5578399 A 14-03-2000 WO 0011178 A 02-03-2000 WO 0011185 A 02-03-2000