Claims
- 1. A method of directing integration of a nucleic acid of interest to a predetermined site, wherein said nucleic acid has homology at or around said predetermined site, in a eukaryote with a preference for nonhomologous recombination, said method comprising:
steering an integration pathway towards homologous recombination.
- 2. The method of directing nucleic acid integration according to claim 1, further comprising: providing a mutant of a component involved in nonhomologous recombination.
- 3. The method of directing nucleic acid integration according to claim 1 or 2, further comprising: inhibiting a component involved in nonhomologous recombination.
- 4. The method according to claim 2 or 3 wherein said component involved in nonhomologous recombination comprises ku70, rad50, mre11, xrs2, lig4 or sir4.
- 5. The method of directing integration of a nucleic acid of interest to a predetermined site according to any one of claims 1 to 3, wherein said nucleic acid of interest is essentially replacing a sequence within said eukaryote.
- 6. The method of directing integration of a nucleic acid of interest to a predetermined site according to claim 5, wherein said component involved in nonhomologous recombination comprises rad50 or xrs2.
- 7. A method of directing integration of a nucleic acid of interest to a subtelomeric region, a telomeric region, or a subtelomeric region and telomeric region in a eukaryote with a preference for nonhomologous recombination by providing a mutant of a component involved in nonhomologous recombination.
- 8. A method of directing integration of a nucleic acid of interest to a subtelomeric region, a telomeric region, or a subtelomeric region and telomeric region in a eukaryote with a preference for nonhomologous recombination, comprising inhibiting a component involved in nonhomologous recombination.
- 9. The method of directing integration according to claim 7 or 8 wherein said component involved in nonhomologous recombination comprises rad50, mre1 or xrs2.
- 10. The method according to any one of claims 1 to 9 wherein said eukaryote is selected from the group consisting of yeast, fungus, and an animal.
- 11. The method according to any one of claims 1 to 10, wherein said nucleic acid of interest is delivered to a cell of said eukaryote by Agrobacterium.
- 12. The method according to any one of claims 1-11 comprising transiently inhibiting integration via nonhomologous recombination.
- 13. The method according claim 12 wherein said transiently inhibiting is provided by an Agrobacterium Vir-fusion protein capable of inhibiting a component involved in nonhomologous recombination.
- 14. The method of directing integration according to claim 13 wherein said Agrobacterium Vir-fusion protein comprises VirF or VirE2.
- 15. The method according to claim 13 or 14 wherein said component involved in nonhomologous recombination comprises ku70, rad50, mre11, xrs2, lig4 or sir4.
- 16. The method according to any one of the foregoing claims wherein said nucleic acid of interest comprises an inactive gene to replace an active gene.
- 17. The method according to any one of claims 1-14, wherein said nucleic acid of interest comprises an active gene to replace an inactive gene.
- 18. The method according to any one of claims 1-14, wherein said nucleic acid of interest encodes a therapeutic proteinaceous substance.
- 19. The method according to any one of claims 1-14, wherein said nucleic acid of interest encodes a substance conferring resistance for an antibiotic substance to a cell.
- 20. The method according to any one of claims 1-14, wherein said nucleic acid of interest confers a desired property to said eukaryote.
- 21. The method according to any one of the foregoing claims wherein said nucleic acid of interest is part of a gene delivery vehicle.
- 22. Use of a method according to any one of claims 1 to 20 for improvement of gene-targeting efficiency.
Priority Claims (1)
Number |
Date |
Country |
Kind |
00204693.6 |
Dec 2000 |
EP |
|
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation of PCT International Patent Application No. PCT/NL/01/00936, filed on Dec. 21, 2001, designating the United States of America, and published, in English, as PCT International Publication No. WO 02/052026 on Jul. 4, 2002, the contents of the entirety of which is incorporated by this reference.
Continuations (1)
|
Number |
Date |
Country |
Parent |
PCT/NL01/00936 |
Dec 2001 |
US |
Child |
10601084 |
Jun 2003 |
US |