Dietary and hormonal regulation of expression of exogenous genes in transgenic animals under control of the promoter of the gene for phosphoenolpyruvate carboxykinase

• US 5,075,229 A
• Filed: 06/13/1988
• Issued: 12/24/1991
• Est. Priority Date: 06/16/1987
• Status: Expired due to Fees

• Current Assignee: MOSAID Technologies Inc. (f/k/a Conversant Intellectual Property Management)
• Original Assignee: Ohio University/Edison Animal Biotechnolgy Center
• Inventors: Short, Jay, McGrane, Mary, Hatzoglou, Maria, Hanson, Richard W.
• Primary Examiner(s): Weimar, Elizabeth C.
• Assistant Examiner(s): Chambers, Jasemine C.
• Application Number:
  US07/205,776
• Time in Patent Office: 1,289 Days
• Field of Search: 800/2, 435/68, 435/172.3, 435/240.2, 435/320, 435/317.1, 935/27, 935/32, 935/53, 935/57, 935/62, 935/111, 514/44
• US Class Current: 514/44.00R
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/20 Animal model comprising regulated expression system
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/02 Animal zootechnically ameliorated
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: C12N 15/00 Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination

Abstract

A transgenic animal is constructed in which one or more cells contain a promoter to the gene for cytosolic PEPCK operably linked to a non-PEPCK gene of interest. Expression of this gene is controlled by modifying the protein and carbohydrate components of the animal'"'"'s diet, or by direct hormonal regulation. The PEPCK promoter is induced by high protein and inhibited by high carbohydrate, or more directly by cAMP and insulin. The linked gene is expressed essentially only after bith and essentially only in particular tissues. The PEPCK promoter has a extremely high promoter strength.

Claims

1. A method of introducing a gene into the liver cells of a mammal in utero which comprises providing a retroviral vector bearing an expressible gene which is foreign to an individual recipient fetal mammal and operably linked to a liver-specific promoter, injecting the vector into the peritoneal cavity of said mammal while it is in a fetal stage of development, and permitting development of said mammal from the fetal stage, during which development the vector is transported to the fetal liver, infects endogenous hemopoietic cells of said fetal liver, and integrates into the genome of said cells conferring on said infected cells the ability to express said gene.

2. The method of claim 1 in which the gene is operably linked to a PEPCK promoter.