Production of membrane proteins in the milk of transgenic mammals

• US 6,743,966 B2
• Filed: 06/27/2002
• Issued: 06/01/2004
• Est. Priority Date: 10/02/1991
• Status: Expired due to Fees

• Current Assignee: Genzyme Corporation (Sanofi )
• Original Assignee: Genzyme Corporation (Sanofi )
• Inventors: Smith, Alan E.
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US10/185,269
• Publication Number:
  US 20030033618A1
• Time in Patent Office: 705 Days
• Field of Search: 800/7, 800/14, 800/15, 800/16, 800/1, 435/69.1, 435/320.1, 435/328, 436/22, 436/23, 554/8
• US Class Current: 800/14
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/10 Mammal
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic

Abstract

Production of proteins not normally secreted through conventional pathways such as membrane proteins including, for example, CFTR associated with cystic fibrosis, is now made possible by collection of such protein from the milk of lactating transgenic animals.

Claims

1. A transgenic non-human mammal which contains within its genome a DNA sequence comprising, as operably linked components, a DNA sequence encoding a membrane protein and a mammary gland specific promoter sequence, wherein, upon expression of the DNA sequence, the membrane protein inserts into an apical membrane of a mammary epithelial cell of the mammal, thereby producing the membrane protein in the milk of the mammal.

2. The mammal of claim 1, wherein the mammary gland specific promoter is a beta-casein promoter.

3. The mammal of claim 1, wherein the mammary gland specific promoter is a beta lactoglobulin promoter.

4. The mammal of claim 1, wherein the mammary gland specific promoter is an alpha-casein promoter.

5. The mammal of claim 1, wherein the mammary gland specific promoter is a whey acid protein promoter.

6. The mammal of claim 1, wherein the membrane protein is a transmembrane ion channel protein.

7. The mammal of claim 1, wherein the membrane protein is selected from the group consisting of a transmembrane ion channel, a hormone receptor protein, a thyroid stimulating hormone receptor, a cytokine receptor protein, and a viral glycoprotein.

8. The mammal of claim 1, wherein the mammal is selected from the group consisting of a mouse, a rabbit, a goat, a sheep and a cow.

9. A transgenic non-human mammal which contains within its genome a DNA construct comprising, as operably linked components, a DNA sequence encoding a membrane protein and a beta casein promoter sequence, wherein, upon expression of said DNA sequence, the membrane protein inserts into an apical membrane of a mammary epithelial cell of the mammal, thereby producing the membrane protein in the milk of the mammal.

10. The mammal of claim 9, wherein the membrane protein is a transmembrane ion channel protein.

11. The mammal of claim 9, wherein the membrane protein is selected from the group consisting of a transmembrane ion channel, a hormone receptor protein, a thyroid stimulating hormone receptor, a cytokine receptor protein, and a viral glycoprotein.

12. The mammal of claim 9, wherein the mammal is selected from the group consisting of a mouse, a rabbit, a goat, a sheep and a cow.

13. A transgenic non-human mammal which contains within its genome a DNA sequence comprising, as operably linked components, a DNA sequence encoding said a cystic fibrosis transmembrane conductance regulator protein and a mammary gland specific promoter sequence, wherein, upon expression of the DNA sequence, the cystic fibrosis transmembrane conductance regulator protein inserts into an apical membrane of mammary epithelial cell of the mammal, thereby producing the cystic fibrosis transmembrane conductance regulator protein in the milk of the mammal.

14. The mammal of claim 13, wherein the mammary specific promoter is a beta-casein promoter.

15. The mammal of claim 13, wherein the mammary gland specific promoter is a beta-lactoglobulin promoter.

16. The mammal of claim 13, wherein the mammary gland specific promoter is an alpha-casein promoter.

17. The mammal of claim 13, wherein said mammary gland specific promoter is a whey acid protein promoter.

18. The mammal of claim 13, wherein the cystic fibrosis transmembrane conductance regulator protein is mature and fully glycosylated.

19. The mammal of claim 13, wherein the mammal is selected from the group consisting of a mouse, a rabbit, a goat, a sheep and a cow.