Claims for Patent: 7,663,024

Summary for Patent: 7,663,024

Title:	P15 hairpin constructs and use
Abstract:	The present invention concerns a method of genetic modification of a TGB-3 wild type viral sequence for reducing or suppressing the possible deleterious effects of the agronomic properties of a transformed plant or plant cell by said TGB-3 viral sequence. The invention further relates to genetically modified TGB-3 viral sequences suitable to induce gene silencing. In particular hairpin constructs based on such sequences proved highly efficient to induce a PTGS mechanism and degradation of the whole of RNA2 thereby. When plants are transformed accordingly the spread of the virus in the plant is significantly reduced or blocked.
Inventor(s):	Lauber; Emmanuelle (Castanet-Tolosan, FR), Guilley; Hubert (Berstett, FR), Richards; Ken (Pfulgriesheim, FR), Jonard; Gerard (Berstett, FR), Klein; Elodie (Strasbourg-Neudorf, FR), Gilmer; David (Strasbourg, FR)
Assignee:	Sesvanderhave, N.V. (Tienen, BE)
Application Number:	11/418,384
Patent Claims:	1. A genetically modified TGB-3 viral sequence comprising a sequence selected from the group consisting of (a) a nucleotide sequence comprising SEQ ID NO: 3 and an antisense sequence of SEQ ID NO: 3; and (b) a nucleotide sequence comprising a modified SEQ ID NO: 3 and an antisense sequence of said modified SEQ ID NO: 3, wherein the modification in SEQ ID NO: 3 consists of a modification that alters a start or a stop codon in SEQ ID NO: 3 so as to inhibit translation, wherein said genetically modified TGB-3 viral sequence when transcribed in a cell is capable of forming a double-stranded self-complementing RNA molecule. 2. The TGB-3 viral sequence of claim 1, wherein the sense and antisense sequences are comprised in one nucleic acid sequence. 3. The TGB-3 viral sequence of claim 2, further comprising an intron fragment interspersed between the sense and antisense sequences, wherein the TGB-3 viral sequence when transcribed in a cell is capable of forming a hairpin RNA molecule. 4. The TGB-3 viral sequence of claim 3, wherein the intron fragment is derived from a plant gene. 5. The TGB-3 viral sequence of claim 4, wherein the plant gene is a beet gene. 6. The TGB-3 viral sequence of claim 3, wherein the intron fragment is an intron fragment of highly transcribed genes. 7. The TGB-3 viral sequence of claim 6, wherein the highly transcribed genes are ribosomal RNA genes. 8. The TGB-3 viral sequence of claim 6, wherein the highly transcribed genes are highly transcribed sugar beet genes. 9. The TGB-3 viral sequence of claim 1 comprising SEQ ID NO: 9. 10. The TGB-3 viral sequence of claim 1 consisting of SEQ ID NO: 9. 11. A vector comprising the genetically modified TGB-3 viral sequence according to claim 1. 12. The vector of claim 11 operably linked to one or more regulatory sequence(s) active in a plant cell. 13. A double stranded self-complementary RNA molecule expressed by a vector of claim 11. 14. A transgenic plant or a transgenic plant cell resistant to a virus and comprising a nucleic acid construct having a genetically modified TGB-3 viral sequence according to claim 1 operably linked to one or more regulatory sequence(s) active in a plant or a plant cell. 15. The transgenic plant or transgenic plant cell according to claim 14, wherein the virus is the BNYVV virus. 16. The transgenic plant or transgenic plant cell according to claim 14, wherein said transgenic plant or transgenic plant cell comprises sugar beet. 17. The transgenic plant or transgenic plant cell according to claim 14, wherein the regulatory sequence comprises a promoter sequence or a terminator sequence that are active in a plant. 18. The transgenic plant of claim 17, wherein said promoter is active in the root tissue of plants. 19. The transgenic plant or transgenic plant cell according to claim 14, wherein said transgenic plant is sugar beet and said transgenic plant cell is a sugar beet cell. 20. The transgenic plant or transgenic plant cell according to claim 14, wherein the regulatory sequence(s) comprise a promoter sequence which is a constitutive or a foreign vegetal promoter sequence. 21. The transgenic plant or transgenic plant cell according to claim 20, wherein the promoter comprises the 35S Cauliflower Mosaic Virus promoter. 22. A transgenic plant tissue derived from the transgenic plant cell according to claim 14, wherein said tissue comprises said genetically modified TGB-3 viral sequence, and wherein said tissue is selected from the group consisting of fruit, stem, root, tuber, and seed. 23. A transgenic reproducible structure obtained from the transgenic plant cell according to claim 14, wherein said reproducible structure comprises said genetically modified TGB-3 viral sequence, and wherein said reproducible structure is selected from the group consisting of calluses, buds, and embryos. 24. A genetically modified TGB-3 viral sequence comprising a sequence selected from the group consisting of: (a) a nucleotide sequence comprising a fragment of SEQ ID NO:3 and an antisense sequence of said fragment of SEQ ID NO:3; and (b) a nucleotide sequence comprising a modified SEQ ID NO: 3 fragment and an antisense sequence of said modified SEQ ID NO: 3 fragment; wherein said fragment of SEQ ID NO:3 is about 25 nucleotides in length with 100% sequence identity to the corresponding part of SEQ ID NO:3, wherein the modification in SEQ ID NO: 3 consists of a modification that alters a start or a stop codon in SEQ ID NO: 3 so as to inhibit translation, wherein said genetically modified TGB-3 viral sequence when transcribed in a cell is capable of forming a double-stranded self-complementing RNA molecule. 25. The TGB-3 viral sequence of claim 24, wherein a nucleic acid sequence comprises the sense and antisense sequences. 26. The TGB-3 viral sequence of claim 25, further comprising an intron fragment interspersed between the sense and antisense sequences, wherein the TGB-3 viral sequence when transcribed in a cell is capable of forming a hairpin RNA molecule. 27. The TGB-3 viral sequence of claim 26, wherein the intron fragment is derived from a plant gene. 28. The TGB-3 viral sequence of claim 27, wherein the plant gene is a beet gene. 29. The TGB-3 viral sequence of claim 26, wherein the intron fragment is an intron fragment of a highly transcribed gene. 30. The TGB-3 viral sequence of claim 29, wherein the highly transcribed gene comprises a ribosomal RNA gene. 31. The TGB-3 viral sequence of claim 29, wherein the highly transcribed gene comprises a highly transcribed sugar beet gene. 32. A vector comprising the genetically modified TGB-3 viral sequence according to claim 24. 33. The vector of claim 32, operably linked to one or more regulatory sequence(s) active in a plant cell. 34. A double stranded self-complementary RNA molecule expressed by a vector of claim 32. 35. A transgenic plant or a transgenic plant cell resistant to a virus and comprising a nucleic acid construct having a genetically modified TGB-3 viral sequence according to claim 24, operably linked to one or more regulatory sequence(s) active in a plant or a plant cell. 36. The transgenic plant or transgenic plant cell according to claim 35, wherein the virus is the BNYVV virus. 37. The transgenic sugar beet plant or transgenic sugar beet plant cell according to claim 35. 38. The transgenic plant or transgenic plant cell according to claim 35, wherein the regulatory sequence comprises a promoter sequence or a terminator sequence. 39. The transgenic plant of claim 38, wherein said promoter is active in the root tissue of a plant. 40. The transgenic plant or transgenic plant cell according to claim 35, wherein said transgenic plant is sugar beet and said transgenic plant cell is a sugar beet cell. 41. The transgenic plant, or transgenic plant cell according to claim 35, wherein the regulatory sequence(s) comprise a promoter sequence which is a constitutive or a foreign vegetal promoter sequence. 42. The transgenic plant or transgenic plant cell according to claim 41, wherein the promoter comprises the 35S Cauliflower Mosaic Virus promoter. 43. A transgenic plant tissue derived from the transgenic plant cell according to claim 35, wherein said tissue comprises said genetically modified TGB-3 viral sequence, and wherein said tissue is selected from the group consisting of fruit, stem, root, tuber, and seed. 44. A transgenic reproducible structure obtained from the transgenic plant cell according to claim 14, wherein said reproducible structure comprises said genetically modified TGB-3 viral sequence, and wherein said reproducible structure is selected from the group consisting of calluses, buds, and embryos. 45. A vector comprising the genetically modified TGB-3 viral sequence according to claim 9. 46. The vector of claim 45 operably linked to one or more regulatory sequence(s) active in a plant cell. 47. A double stranded self-complementary RNA molecule expressed by a vector of claim 45. 48. A transgenic plant or a transgenic plant cell resistant to a virus and comprising a nucleic acid construct having a genetically modified TGB-3 viral sequence according to claim 9 operably linked to one or more regulatory sequence(s) active in a plant or a plant cell. 49. The transgenic plant or transgenic plant cell according to claim 48, wherein the virus is the BNYVV virus. 50. The transgenic plant or transgenic plant cell according to claim 48, wherein said transgenic plant or transgenic plant cell comprises sugar beet. 51. The transgenic plant or transgenic plant cell according to claim 48, wherein the regulatory sequence comprises a promoter sequence or a terminator sequence that is active in a plant. 52. The transgenic plant or transgenic plant cell according to claim 48, wherein said transgenic plant is sugar beet and said transgenic plant cell is a sugar beet cell. 53. The transgenic plant or transgenic plant cell according to claim 48, wherein the regulatory sequence(s) comprise a promoter sequence which is a constitutive or a foreign vegetal promoter sequence. 54. The transgenic plant or transgenic plant cell according to claim 53, wherein the promoter comprises the 35S Cauliflower Mosaic Virus promoter. 55. A transgenic plant tissue comprising the genetically modified TGB-3 sequence of claim 9, wherein said tissue is derived from the transgenic plant cell according to claim 48, and wherein said tissue is selected from the group consisting of fruit, stem, root, tuber, and seed. 56. A transgenic reproducible structure comprising a genetically modified TGB-3 sequence of claim 9, wherein said genetically modified TGB-3 viral sequence when transcribed in a cell is capable of forming a double-stranded self-complementing RNA molecule, wherein said reproducible structure is obtained from the transgenic plant cell according to claim 48, and wherein said reproducible structure is selected from the group consisting of calluses, buds, and embryos. 57. A vector comprising the genetically modified TGB-3 viral sequence according to claim 10. 58. A nucleotide sequence comprising a modified SEQ ID NO: 3 and an antisense sequence of said modified SEQ ID NO: 3, wherein the modifications in SEQ ID NO: 3 consist of a modification which alters a start codon in SEQ ID NO: 3 so as to inhibit translation and a modification which alters a stop codon in SEQ ID NO: 3 so as to inhibit translation, wherein the nucleotide sequence when transcribed in a cell is capable of forming a double-stranded self-complementing RNA molecule.

Details for Patent 7,663,024

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2039-02-26
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2039-02-26
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2039-02-26