| US 7,579,326 B2 | ||
| Gene switch systems employing regulators with decreased dimerization | ||
| Ronald V Abruzzese, The Woodlands, Tex. (US); Vidya Mehta, Houston, Tex. (US); Jeffrey L Nordstrom, College Station, Tex. (US); Jason Fewell, The Woodlands, Tex. (US); Bert O'Malley, Houston, Tex. (US); and Sophia Tsai, Houston, Tex. (US) | ||
| Assigned to Genetronics, Inc., San Diego, Calif. (US); and Baylor College of Medicine, Houston, Tex. (US) | ||
| Filed on Mar. 25, 2003, as Appl. No. 10/400,053. | ||
| Application 10/400053 is a continuation of application No. PCT/US01/30305, filed on Sep. 25, 2001. | ||
| Claims priority of provisional application 60/278281, filed on Mar. 23, 2001. | ||
| Claims priority of provisional application 60/260781, filed on Jan. 10, 2001. | ||
| Claims priority of provisional application 60/235030, filed on Sep. 25, 2000. | ||
| Prior Publication US 2003/0220286 A1, Nov. 27, 2003 | ||
| Int. Cl. A01N 43/04 (2006.01); A61K 31/70 (2006.01); A01N 63/00 (2006.01); A01N 65/00 (2006.01); C12N 15/00 (2006.01); C07H 19/00 (2006.01); C07H 21/00 (2006.01); C07H 21/04 (2006.01) | ||
| U.S. Cl. 514—44 [424/93.1; 435/320.1; 536/23.1; 536/23.4; 536/24.5; 536/24.1] | 50 Claims |
| 1. An inducible expression system comprising:
(a) a first expression cassette including a promoter operably linked to a first nucleic acid sequence encoding a regulator
protein, wherein the regulator protein comprises:
a mutated GAL-4 DNA-binding domain that carries a mutation in a helical domain located in a region from amino acid 54-93 of
SEQ ID NO: 10, wherein the mutation in the helical domain decreases dimerization of the regulator protein occurring in the
absence of a anti-progestin ligand,
a transregulatory domain selected from the group consisting of NFkappaBp65, VP-16, TAF-1, TAF-2, TAU-1, TAU-2, TEF-1 and ORF-10
transregulatory domains, and
a ligand-binding domain of a progesterone receptor, wherein the ligand binding domain has a deletion in naturally occurring
amino acids in a region encompassing from about 1 to about 60 carboxyl terminal amino acids of the ligand binding domain,
said deletion conferring an ability to be activated by an anti-progestin ligand; and
(b) a second expression cassette having a second nucleic acid sequence encoding a desired product, operably linked to a promoter
comprising a GAL-4 binding site, wherein the regulator protein is activated in the presence of the anti-progestin and the
expression of the second nucleic acid sequence is controlled by binding of the activated regulator protein to Gal-4 binding
sites in the promoter region of the second expression cassette.
|