Expression of active human factor IX in mammary tissue and of milk non human transgenic mammals

Recombinant Factor IX characterized by a high percentage of active protein can be obtained in the milk of transgenic animals that incorporate chimeric DNA molecules according to the present invention. Transgenic animals of the present invention are produced by introducing into developing embryos DNA that encodes Factor IX, such that the foreign DNA is stably incorporated in the DNA of germ line cells of the mature animal. Particularly efficient expression was accomplished using a chimeric construct comprising a mammary gland specific promoter, Factor IX cDNA that lacked the complete or any portion of the 5'untranslated and 3'-untranslated region, which is substituted with a 5'and 3'-end of the mouse whey acidic protein gene. In vitro cell cultures of cells explanted from the transgenic mammal of the invention and methods of producing Factor IX from such said culture and methods of treating hemophilia B are also described. Inventors: Velander; William H. (Blacksburg, VA); Drohan; William N. (Springfield, VA); Lubon; Henryk (Rockville, MD); Johnson; John L. (late of Blacksburg, VA) Assignee: American Red Cross (Rockville, MD); Virginia Tech Intellectual Properties, Inc. (Blacksburg, VA) Appl. No.: 367087 Filed: September 15, 1999 PCT Filed: February 13, 1998 PCT NO: PCT/US98/02638 371 Date: March 1, 2000 102(e) Date: March 1, 2000 PCT PUB.NO.: WO98/35689 PCT PUB. Date: August 20, 1998 Current U.S. Class: 800/14; 435/69.6; 435/325; 800/7; 800/15; 800/16; 800/17; 800/18 Intern'l Class: A01K 067/027 Field of Search: 800/7,14,15,16,17,18 435/325,69.6 References Cited [Referenced By] U.S. Patent Documents 4786726 Nov., 1988 Smith. 4873316 Oct., 1989 Meade et al. 5171569 Dec., 1992 Anson et al. 5322775 Jun., 1994 Clark et al. 5366894 Nov., 1994 Clark et al. 5476995 Dec., 1995 Clark et al. 5589604 Dec., 1996 Drohan et al. 5831141 Nov., 1998 Lubon et al. 800/2. Foreign Patent Documents 0 791 652 Aug., 1997 EP. 0 274 489 Oct., 1997 EP. 0 396 699 Oct., 1997 EP. 90/05188 May., 1990 WO. 91/08216 Jun., 1991 WO. 94/05796 Mar., 1994 WO. 95/30000 Nov., 1995 WO.