Complex

• US 20040214303A1
• Filed: 01/13/2003
• Published: 10/28/2004
• Est. Priority Date: 09/20/2000
• Status: Abandoned Application

• Current Assignee: Takara Bio Incorporated (Ningbo Junsheng Investment Group Co., Ltd.)
• Original Assignee: Takara Bio Incorporated (Ningbo Junsheng Investment Group Co., Ltd.)
• Inventors: Chono, Hideto, Ueno, Mitsuhiro, Matsumura, Hajime, Kato, Ikunoshin
• Application Number:
  US10/332,704
• Publication Number:
  US 20040214303A1
• Time in Patent Office: Days
• US Class Current: 435/252.3
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 15/90 Stable introduction of foreign DNA into chromosome
• CPC Class Codes: C12N 15/907 in mammalian cells
• CPC Class Codes: C12N 2740/13022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2740/13043 viral genome or elements thereof as genetic vector

Abstract

A complex comprising a double-stranded DNA containing a terminal repeated sequence originating in a retrovirus and a DNA having a base sequence which does not occur in this retrovirus and a retrovirus-origin protein component.

Claims

1. A complex which comprises a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus as well as a protein component derived from a retrovirus.

2. The complex according to claim 1, wherein the double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus is derived from a recombinant retrovirus.

3. The complex according to claim 2, wherein the recombinant retrovirus is obtainable by infecting a cell susceptible to the recombinant retrovirus with the recombinant retrovirus.

4. The complex according to claim 3, wherein the cell cycle of the cell susceptible to the recombinant retrovirus is arrested.

5. The complex according to claim 1, which is obtainable by mixing an isolated double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus with an isolated protein component derived from a retrovirus.

6. The complex according to claim 1, wherein the protein component derived from a retrovirus is derived from a culture supernatant of a packaging cell and/or from an extract of a packaging cell.

7. The complex according to claim 1, wherein the protein component derived from a retrovirus contains an enzyme having an activity of integrating a DNA having a long terminal repeat derived from a retrovirus into a chromosomal DNA.

8. A method for producing a complex, the method comprising:

(a) infecting a cell susceptible to a recombinant retrovirus with the recombinant retrovirus that contains a long terminal repeat derived from a retrovirus and a nucleotide sequence that is absent in the retrovirus;

(b) culturing the cell infected in step (a); and

(c) collecting a complex that comprises a double-stranded DNA derived from the recombinant retrovirus and a protein component derived from the retrovirus from the cell cultured in step (b).

9. The method according to claim 8, wherein the cell cycle of the cell susceptible to a recombinant retrovirus is arrested.

10. A method for producing a complex, the method comprising:

(a) destroying a particle of a recombinant retrovirus that contains a long terminal repeat derived from a retrovirus and a nucleotide sequence that is absent in the retrovirus to prepare a mixture containing a viral component;

(b) preparing a double-stranded DNA derived from the recombinant retrovirus by adding deoxyribonucleotides to the mixture prepared in step (a) to form a complex that comprises the double-stranded DNA and a protein component derived from a retrovirus; and

(c) collecting the complex formed in step (b).

13. The method according to any one of claims 8, 10 and 11, wherein the protein component derived from a retrovirus contains an enzyme having an activity of integrating a DNA having a long terminal repeat derived from a retrovirus into a chromosomal DNA.

15. The method according to claim 14, wherein the complex is obtainable by the method defined by any one of claims 8, 10 and 11.

21. The composition according to claim 20, wherein the complex is obtainable by the method defined by any one of claims 8, 10 and 11.

34. The method according to claim 33, wherein the complex is obtainable by the method defined by any one of claims 8, 10 and 11.

40. The composition according to claim 38, wherein the complex is obtainable by the method defined by any one of claims 8, 10 and 11.

11. A method for producing a complex, the method comprising:

(a) isolating a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus;

(b) isolating a protein component derived from a retrovirus;

(c) mixing the double-stranded DNA and the protein component derived from a retrovirus obtained in the previous steps together to form a complex; and

(d) collecting the complex formed in the previous step.

12. The method according to claim 11, wherein the protein component derived from a retrovirus is a culture supernatant of a packaging cell and/or an extract of a packaging cell.

14. A method for transferring a gene into a cell, the method comprising:

transferring, into a cell, a complex that comprises a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus as well as a protein component derived from a retrovirus.

52. The method according to claim 14 or 16, wherein the gene is transferred into a eukaryotic cell.

53. The method according to claim 14 or 16, wherein the gene is transferred into a cultured cell derived from a eukaryotic cell.

54. The method according to claim 14 or 16, wherein the gene is transferred into a eukaryotic individual.

55. The method according to claim 14 or 16, wherein the gene is transferred into a vertebrate fertilized egg.

56. The method according to claim 14 or 16, wherein the gene is transferred into a vertebrate germ line.

16. A method for transferring a gene into a cell, the method comprising:

transferring, into a cell, a protein component derived from a retrovirus as well as a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus.

17. The method according to claim 16, wherein the protein component derived from a retrovirus is derived from a culture supernatant of a packaging cell.

18. The method according to claim 16, wherein the protein component derived from a retrovirus contains an enzyme having an activity of integrating a DNA having a long terminal repeat derived from a retrovirus into a chromosomal DNA.

19. A composition for transferring a gene into a cell, which contains a protein component derived from a retrovirus.

20. The composition according to claim 19, wherein the protein component derived from a retrovirus is forming a complex with a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus.

22. The composition according to claim 19, which further contains a vector having a long terminal repeat derived from a retrovirus.

23. The composition according to claim 22, wherein the vector is a vector for preparing a recombinant retrovirus.

24. The composition according to claim 22, wherein the vector is a plasmid vector.

25. The composition according to claim 19, wherein the protein component derived from a retrovirus is derived from a culture supernatant of a packaging cell and/or from an extract of a packaging cell.

26. The composition according to claim 19, wherein the protein component derived from a retrovirus contains an enzyme having an activity of integrating a DNA having a long terminal repeat derived from a retrovirus into a chromosomal DNA.

27. A kit for transferring a gene into a cell, which contains a composition for transferring a gene into a cell that contains a protein component derived from a retrovirus.

28. A kit for transferring a gene into a cell,

which is used for a method for transferring a gene into a cell,

which is in a packed form, and

which contains instructions that direct transferring, into a cell, of a complex that comprises a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus as well as a protein component derived from a retrovirus.

29. A kit for transferring a gene into a cell,

which is used for a method for transferring a gene into a cell,

which is in a packed form, and

which contains instructions that direct transferring, into a cell, of a protein component derived from a retrovirus as well as a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus.

30. A product of a reagent for transferring a gene into a cell,

which contains a packing material and a reagent for gene transfer enclosed in the packing material,

wherein the reagent for gene transfer contains a complex that comprises a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus as well as a protein component derived from a retrovirus and/or a reagent for preparing the complex, and

wherein it is indicated in a label stuck to the packing material or instructions attached to the packing material that the reagent for gene transfer can be used for gene transfer that does not comprise a step of infecting a cell of interest with a recombinant retrovirus.

32. The product according to claim 30 or 31, wherein the protein component derived from a retrovirus contains an enzyme having an activity of integrating a DNA having a long terminal repeat derived from a retrovirus into a chromosomal DNA.

31. A product of a reagent for transferring a gene into a cell,

which contains a packing material and a reagent for gene transfer enclosed in the packing material,

wherein the reagent for gene transfer contains a protein component derived from a retrovirus, and

wherein it is indicated in a label stuck to the packing material or instructions attached to the packing material that the reagent for gene transfer can be used for gene transfer that does not comprise a step of infecting a cell of interest with a recombinant retrovirus.

33. A method for transducing a cell, the method comprising:

transferring, into a cell, a complex that comprises a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus as well as a protein component derived from a retrovirus.

57. The method according to claim 33 or 35, wherein a eukaryotic cell is transduced.

58. The method according to claim 33 or 35, wherein a cultured cell derived from a eukaryotic cell is transduced.

59. The method according to claim 33 or 35, wherein a eukaryotic individual is transduced.

60. The method according to claim 33 or 35, wherein a vertebrate fertilized egg is transduced.

61. The method according to claim 33 or 35, wherein a vertebrate germ line is transduced.

35. A method for transducing a cell, the method comprising:

transferring, into a cell, a protein component derived from a retrovirus as well as a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus.

36. The method according to claim 35, wherein the protein component derived from a retrovirus is derived from a culture supernatant of a packaging cell and/or from an extract of a packaging cell.

37. The method according to claim 35, wherein the protein component derived from a retrovirus contains an enzyme having an activity of integrating a DNA having a long terminal repeat derived from a retrovirus into a chromosomal DNA.

38. A composition for transducing a cell, which contains a protein component derived from a retrovirus.

39. The composition according to claim 38, wherein the protein component derived from a retrovirus is forming a complex with a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus.

41. The composition according to claim 38, which further contains a vector for preparing a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus.

42. The composition according to claim 41, wherein the vector is a vector for preparing a recombinant retrovirus.

43. The composition according to claim 41, wherein the vector is a plasmid vector.

44. The composition according to claim 38, wherein the protein component derived from a retrovirus is derived from a culture supernatant of a packaging cell and/or from an extract of a packaging cell.

45. The composition according to claim 38, wherein the protein component derived from a retrovirus contains an enzyme having an activity of integrating a DNA having a long terminal repeat derived from a retrovirus into a chromosomal DNA.

46. A kit for transducing a cell, which contains a composition for transducing a cell that contains a protein component derived from a retrovirus.

47. A kit for transducing a cell, which contains instructions that direct transferring, into a cell, of a complex that comprises a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus as well as a protein component derived from a retrovirus for a method for transducing a cell.

48. A kit for transducing a cell,

which is used for a method for transducing a cell,

which is in a packed form, and

which contains instructions that direct transferring, into a cell, of a protein component derived from a retrovirus as well as a double-stranded DNA containing a long terminal repeat derived from a retrovirus and a DNA having a nucleotide sequence that is absent in the retrovirus.

49. A product of a reagent for transducing a cell,

which contains a packing material and a reagent for transduction enclosed in the packing material,

wherein the reagent for transduction contains a complex that comprises a double-stranded DNA derived from a recombinant retrovirus and a protein component derived from a retrovirus and/or a reagent for preparing the complex, and

wherein it is indicated in a label stuck to the packing material or instructions attached to the packing material that the reagent for transduction can be used for transduction that does not comprise a step of infecting a cell of interest with a recombinant retrovirus.

51. The product according to claim 49 or 50, wherein the protein component derived from a retrovirus is an enzyme having an activity of integrating a DNA having a long terminal repeat derived from a retrovirus into a chromosomal DNA.

50. A product of a reagent for transducing a cell,

which contains a packing material and a reagent for transduction enclosed in the packing material,

wherein the reagent for transduction contains a protein component derived from a retrovirus, and

wherein it is indicated in a label stuck to the packing material or instructions attached to the packing material that the reagent for transduction can be used for transduction that does not comprise a step of infecting a cell of interest with a recombinant retrovirus.