Transgenic animals secreting desired proteins into milk

• US 6,727,405 B1
• Filed: 05/19/1994
• Issued: 04/27/2004
• Est. Priority Date: 04/09/1986
• Status: Expired due to Term

• Current Assignee: Genzyme Corporation (Sanofi ), The United States of America As Represented By The Secretary of Agriculture
• Original Assignee: Genzyme Corporation (Sanofi )
• Inventors: Gordon, Katherine, Groet, Suzanne, Westphal, Heiner, Hennighausen, Lothar
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US08/246,259
• Time in Patent Office: 3,631 Days
• Field of Search: 800/2, 800/7, 800/14, 800/15, 800/16, 800/17, 800/18, 536/23.1, 536/24.1, 536/23.5, 536/23.51, 435/69.1, 435/69.2, 435/69.7, 435/71.1, 435/172.3, 435/320.1
• US Class Current: 800/14
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/10 Mammal
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 15/89 using microinjection
• CPC Class Codes: C12N 2730/10122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: C12N 2830/85 mammalian
• CPC Class Codes: C12N 9/6459 t-plasminogen activator (3.4.21.68), i.e. tPA
• CPC Class Codes: C12Y 304/21069 Protein C activated (3.4.21.69)

Abstract

A DNA sequence containing a gene encoding a protein, the gene being under the transcriptional control in the DNA sequence of a mammalian milk protein promoter which does not naturally control the transcription of the gene, such DNA sequence including DNA enabling secretion of the protein.

Claims

1. A non-human mammal whose genome comprises a DNA construct comprising a whey acidic protein promoter operably linked to a DNA sequence encoding a heterologous protein, wherein said construct further comprises a DNA sequence encoding a secretory peptide operatively linked to said DNA sequence encoding a heterologous protein, wherein said mammal is selected from the group consisting of mouse, sheep, pig, goat and cow, and wherein said heterologous protein is expressed in the milk of the mammal.

2. A method of producing a protein comprising the steps of:

(a) providing an adult, female non-human mammal whose genome comprises a DNA construct comprising a whey acidic protein promoter operably linked to a DNA sequence encoding a heterologous protein, wherein said construct further comprises a DNA sequence encoding a secretory peptide operatively linked to said DNA sequence encoding a heterologous protein, wherein said mammal is selected from the group consisting of mouse, sheep, pig, goat and cow,

(b) inducing lactation in said mammal, thereafter

(c) collecting milk from said lactating mammal, and

(d) isolating said heterologous protein from said collected milk.

3. The method of claim 2, wherein the mammal of step (a) was developed from a fertilized ovum comprising said DNA construct.

4. The method of claim 2, wherein said protein is biologically active human tissue plasminogen activator.

5. A DNA construct comprising (i) a whey acidic protein promoter operably linked to a DNA sequence encoding a heterologous protein, and wherein said construct further comprises (ii) a DNA sequence encoding a secretory peptide operatively linked to said DNA sequence encoding (iii) a heterologous protein.