FLP-mediated gene modification in mammalian cells, and...

Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...

Reexamination Certificate

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C435S325000

Reexamination Certificate

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07371577

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 fromSaccharomyces. 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 β-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 vertebrate development. The FLP recombination system of the present invention can be incorporated in transgenic, non-human mammals to achieve site-specific integration of transgenes, to construct functional genes or to disrupt existing genes.

REFERENCES:
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Golic, 1989, Cell, vol. 59, pp. 499-509.
Dymecki, 1996, PNAS, vol. 93, pp. 6191-6196.
Le Mouellic, 1990, PNAS, vol. 87, pp. 4712-4716.
Awatramani, 2001, Nature Gnetics, vol. 29, pp. 257-259.
Soriano, 1999, Nature Gnetics, vol. 21, pp. 70-71.
Mullins, 1990; Nature, vol. 344, pp. 541-544.
Hammer, 1990, Cell, vol. 63, 1099-1112.
Mullins, 1989, EMBO J., vol. 8, pp. 4065-4072.
Taurog, 1988, Jour. Immunol., vol. 141, pp. 4020-4023.
Wall, 1996, Theriogenology, vol. 45, pp. 57-68.
Overbeek, 1994, “Factors affecting transgenic animal production,” Transgenic animal technology, pp. 96-98).
Mullins, 1996, J. Clin. Invest., vol. 98, pp. S37-S40.
Golic, 1989, Cell, vol. 59, pp. 499-509.
Dymecki, 1996, PNAS, vol. 93, pp. 6191-6196.
Le Mouellic, 1990, PNAS, vol. 87, pp. 4712-4716.
Awatramani, 2001, Nature Gnetics, vol. 29, pp. 257-259.
Soriano, 1999, Nature Gnetics, vol. 21, pp. 70-71.

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