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

1995-06-07

1997-10-14

Low, Christopher S. F.

Chemistry: molecular biology and microbiology

Animal cell, per se ; composition thereof; process of...

4351723, 4353201, 435350, 435351, 435352, 435353, 435354, 435363, 435366, 435193, 435197, 435194, 536 242, 536 241, 536 2374, 536 231, 935 66, 935 8, 935 22, 514 44, 800 2, C12N 506, C12N 1509, C12N 1590

Patent

active

056771770

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.

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