FLP-mediated gene modification in mammalian cells, and compositi

Details FLP-mediated gene modification in mammalian cells, and compositi FLP-mediated gene modification in mammalian cells, and compositi

1997-04-08

1999-03-23

Low, Christopher S. F.

Chemistry: molecular biology and microbiology

Process of mutation, cell fusion, or genetic modification

Introduction of a polynucleotide molecule into or...

435325, 435462, 435463, 435466, 435468, 435471, 4353201, 435350, 435351, 435352, 435353, 435354, 435363, 435366, 435193, 435197, 435194, 536 242, 536 241, 536 2374, 536 231, 514 44, 800 8, 800 3, 935 8, 935 66, 935 22, 935 44, C12N 1563, C12N 1587

Patent

active

058858369

ABSTRACT:
A gene activation/inactivation and site-specific integration system has been developed for mammalian cells. The invention system is based on the recombination of transfected sequences by FLP, a recombinase derived from Saccharomyces. In several cell lines, FLP has been shown to rapidly and precisely recombine copies of its specific target sequence. For example, a chromosomally integrated, silent .beta.-galactosidase reporter gene was activated for expression by FLP-mediated removal of intervening sequences to generate clones of marked cells. Alternatively, the reverse reaction can be used to target transfected DNA to specific chromosomal sites. These results demonstrate that FLP can be used, for example, to mosaically activate or inactivate transgenes for a variety of therapeutic purposes, as well as for analysis of vertebriate development.

REFERENCES:
patent: 5654182 (1997-08-01), Wahl et al.
patent: 5677177 (1997-10-01), Wahl et al.
Wall (1996) Theriogenology 45: 57-68.
Houdebine (1994) J. Biotechnol. 34: 269-287.
O'Gorman et al. (1991) Science 251: 1351-1355.
Aladjem et al. (1997) Molec. Cell. Biol. 17(2) 857-861.

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