Prior Art Request for US20140296324 A1 (Virovek Incorporation) —Vectors harboring toxic genes, methods and uses therefor

Question

This application from VIROVEK INCORPORATION seeks to patent a method of healing cancer! 10 minutes of your time can help narrow this US patent application before it becomes a patent.

QUESTION - Have you seen anything that was published before 2013/08/15 that discusses:

• The use of a insect cell for a baculovirus, a retrovirus, an adenovirus, an adeno-associated virus production.
• The use of a toxic gene for mammals not expressed in insect cells.

If you've ever seen anything like this before, please submit evidence of prior art as an answer to this question.

EXTRA CREDIT - Include a reference to anything that meets all of the criteria to the question above AND ALSO the use of a gene in a viral vector expressed in a insect cell or method of obtaining it.

TITLE: VECTORS HARBORING TOXIC GENES, METHODS AND USES THEREFOR

Summary: The present disclosure describes nucleic acids, and viruses comprising such nucleic acids, for growing a toxic gene in an insect cell. These nucleic acids comprise a sequence encoding a toxic polypeptide, and an intron that interrupts the sequence, whereby the intron is spliced in mammalian cells but not in insect cells. Infection of mammalian cells but not insect cells with the nucleic acids or viruses can lead to expression of toxic levels of the toxic polypeptide in mammalian cells but not in insect cells. Viruses, such as an AAV or a baculovirus comprising a nucleic acid can be grown in insect cell lines in vitro and can be administered to a subject in need of therapy, such as a subject in need of cancer therapy.

• Publication Number: US2014296324

• Assignee: CHEN HAIFENG; VIROVEK INC

• Prior Art Date: Seeking prior Art predating 16 Oct. 2012
• Open for Challenge at USPTO: Open through 2 Apr. 2015

The claims verbatim:

1. A nucleic acid comprising: a sequence encoding a toxic polypeptide; and an intron that interrupts the sequence, whereby the intron is spliced in mammalian cells but not in insect cells to form an mRNA that is translated to form cell-toxic levels of the toxic polypeptide in mammalian cells but not in insect cells.
2. A nucleic acid in accordance with claim 1, wherein the toxic polypeptide is selected from the group consisting of diphtheria toxin (DT-A), barnase, ricin, abrin, and Pseudomonas exotoxin.
3. A nucleic acid in accordance with claim 1, wherein the toxic polypeptide is diphtheria toxin (DT-A).
4. A nucleic acid in accordance with claim 1, wherein the toxic polypeptide is a barnase polypeptide.
5. A nucleic acid in accordance with claim 1, further comprising at least one expression element operably linked to the sequence.
6. A nucleic acid in accordance with claim 5, wherein the expression element is selected from the group consisting of a promoter, an IRES, an enhancer and a combination thereof.
7. A nucleic acid in accordance with claim 5, wherein the expression element is selected from the group consisting of a CMV promoter, an AFP promoter, an AFP enhancer, an hSURV promoter, a CXCR4 promoter, an hTERT promoter, a COX2 promoter, and a CCKAR promoter.
8. A nucleic acid in accordance with claim 5, wherein the expression element is selected from the group consisting of an hSURV promoter and a CXCR4 promoter.
9. A nucleic acid in accordance with any one of claims 1-8, wherein the intron is an artificial intron.
10. A nucleic acid in accordance with any one of claims 1-8, wherein, the intron is a human growth hormone intron.
11. A nucleic acid in accordance with any one of claims 1-8, wherein the intron is a SV40 large T-antigen intron.
12. A nucleic acid in accordance with any one of claims 1-8, comprising, in 5′ to 3′ order, a first ITR, a promoter, a first portion of a toxic gene, an intron, a second portion of the toxic gene, a polyadenylation signal, and a second ITR.
13. A nucleic acid in accordance with claim 13, wherein the first ITR is an AAV ITR.
14. A nucleic acid in accordance with claim 13, wherein the second ITR is an AAV ITR.
15. A nucleic acid in accordance with claim 13, wherein the promoter is a CMV promoter.
16. A nucleic acid in accordance with claim 13, wherein the promoter is a hSURV promoter.
17. A nucleic acid in accordance with claim 13, wherein the promoter is a hCXCR4 promoter.
18. A nucleic acid in accordance with claim 13, wherein the promoter is a hAFP promoter.
19. A nucleic acid in accordance with claim 13, wherein the toxin gene encodes DT-A.
20. A nucleic acid in accordance with claim 13, wherein the toxic gene encodes Barnase.
21. A viral vector comprising the nucleic acid of any one of claims 1-8.
22. A viral vector in accordance with claim 22, wherein the vector is selected from the group consisting of a baculovirus, a retrovirus, an adenovirus, an adeno-associated virus (AAV) and a combination thereof.
23. A viral vector in accordance with claim 22, wherein the vector is a baculovirus, an adeno-associated virus (AAV), or a combination thereof.
24. A viral vector in accordance with claim 22, wherein the vector is an adeno-associated virus (AAV).
25. A viral vector in accordance with claim 22, wherein the vector is an AAV2.
26. A viral vector in accordance with claim 22, wherein the vector is an AAV9.
27. A viral vector in accordance with claim 22, wherein the vector comprises an ITR.
28. An insect cell in vitro comprising the nucleic acid of any one of claims 1-8.
29. An insect cell in accordance with claim 29, wherein the cell is selected from the group consisting of a Trichoplusia ni BTI-Tn-5B1-4 cell, a Spodoptera frugiperda Sf9 cell and a Spodoptera frugiperda Sf21 cell.
30. An insect cell in accordance with claim 29, wherein the nucleic acid is comprised by a viral genome.
31. An insect cell in accordance with claim 29, wherein the nucleic acid is comprised by a virus.
32. An insect cell in accordance with claim 29, wherein the cell is stably transformed with the nucleic acid of any one of claims 1-8.
33. A cell culture comprising: a plurality of cells of claim 29; and a culture medium.
34. A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises greater than 109 viral genomes/ml.
35. A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises greater than 1010 viral genomes/ml.
36. A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises greater than 1011 viral genomes/ml.
37. A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises greater than 1012 viral genomes/ml.
38. A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises a baculovirus greater than 106 PFU/ml.
39. A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises a baculovirus greater than 107 PFU/ml.
40. A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises a baculovirus greater than 108 PFU/ml.
41. A cell culture in accordance with claim 34, wherein unconcentrated culture medium comprises a baculovirus greater than 109 PFU/ml.
42. A method of growing a vector comprising a toxic gene in vitro, comprising: providing a cell culture comprising insect cells; infecting or transfecting the cells with a nucleic acid of any one of claims 1-8; and incubating the cells under conditions suitable for virus production.
43. A method of treating a cancer, comprising administering to a subject in need of therapy a therapeutically effective amount of the nucleic acid of any one of claims 1-8.
44. A method of treating a cancer, comprising administering a therapeutically effective amount of an adeno-associated virus comprising the nucleic acid of any one of claims 1-8 to a subject in need of therapy.
45. A method of treating a cancer in accordance with claim 45 wherein the cancer is selected from the group consisting of prostate cancer, breast cancer, brain tumor, cervical cancer and nasopharyngeal carcinoma.
46. Use of a nucleic acid of any one of claims 1-8 for the treatment of a cancer.
47. A nucleic acid in accordance with any one of claims 1-8 for use in the treatment of a cancer.
48. Use of a nucleic acid of any one of claims 1-8 in the manufacture of a medicament for treatment of a cancer.
49. Use of a viral vector of claim 22 for the treatment of a cancer.
50. The viral vector of claim 22 for use in the treatment of a cancer.
51. Use of a viral vector of claim 22 in the manufacture of a medicament for treatment of a cancer.
52. A method of producing AAV comprising a toxic gene in vitro, comprising: providing a cell culture comprising insect cells; infecting the cells with a virus vector of claim 24 and with Bac-inCap-inRep for at least about 1 day; and lysing the cells to form a lysate comprising the AAV.
53. A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for at least 2 days.
54. A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for at least 3 days.
55. A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for about 3 days.
56. A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for at least 4 days.
57. A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for at least 5 days.
58. A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for up to about 5 days.
59. A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for up to about 6 days.
60. A method in accordance with claim 53, wherein the infecting the cells for at least about 1 day comprises infecting the cells for up to about 7 days.
61. A method in accordance with claim 53, further comprising digesting the cellular debris with benzonase.
62. A method in accordance with claim 53, wherein the AAV is AAV2.
63. A method in accordance with claim 53, wherein the AAV is AAV9.
64. A method in accordance with claim 53, wherein the insect cells are selected from the group consisting of Trichoplusia ni BTI-Tn-5B1-4 cells, Spodoptera frugiperda Sf9 cells, Spodoptera frugiperda Sf21 cells, and any combination thereof.
65. A method in accordance with claim 53, wherein the insect cells are Spodoptera frugiperda Sf9 cells.

Here there is an image how the claims are related (from the espacenet web):

I think many claims might are invalidated. I doubt these system has never been thought before. Or the terminology is not clear enough, what does here the toxic gene mean? My concern is about using some of the information from these patent in other diseases

Note: If I can improve my question, please let me know.