Gene therapy using gene fusions for genetic or acquired disorders

• US 5,166,059 A
• Filed: 05/03/1991
• Issued: 11/24/1992
• Est. Priority Date: 06/16/1987
• Status: Expired due to Term

• Current Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Original Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Inventors: Gottesman, Michael, Pastan, Ira
• Primary Examiner(s): Ellis, Joan
• Application Number:
  US07/697,000
• Time in Patent Office: 571 Days
• Field of Search: 435/69.1, 435/69.7, 435/91, 435/172.3, 435/235.1, 435/320.1, 435/240.2, 536/27, 570/350
• US Class Current: 435/69.7
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• 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 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2799/027 where the vector is derived from a retrovirus
• CPC Class Codes: C12N 9/78 acting on carbon to nitrogen bonds other than peptide bonds (3.5)

Abstract

Gene therapy utilizing an MDR1 linked fusion coding sequence has been disclosed. ADA activity has been introduced into cells using MDR1 linked fusion gene.

Claims

1. A nucleotide sequence encoding an MDR1-ADA fusion gene comprising the coding sequence for the human MDR1 gene directly linked to the 5'"'"' end of the coding sequence for the ADA gene, wherein the linked coding sequences form a single open reading frame.

2. An expression vector comprising the fusion gene of claim 1.

4. The expression vector of claim 2, wherein said expression vector is a retroviral vector.

5. The expression vector of claim 4 wherein said vector further comprises the Harvey sarcoma viral LTR promoter operably linked to the nucleotide sequence encoding the MDR1-ADA fusion protein.

6. A mammalian cell transfected with the vector of claim 2.

7. A method for expression the MDR1-ADA fusion gene which comprises culturing the mammalian cells of claim 6 under suitable conditions so that the MDR1-ADA fusion protein is produced by said cells.

8. The method of claim 7 wherein said cells are cultured in the presence of a stimulatory drug under conditions that result in amplification of the MDR1-ADA fusion protein.

3. An expression vector having the identifying characteristics of ATCC 67699.