Method for the stable inversion of DNA sequence by...

Chemistry: molecular biology and microbiology – Vector – per se

Reexamination Certificate

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C536S023100

Reexamination Certificate

active

07074611

ABSTRACT:
The invention relates to a method for the stable inversion of a DNA fragment upon recombinase-mediated rearrangements using two sets of two incompatible site-specific recombinase targeting sites (SSRTS) in the same order but in reverse orientation flanking the DNA fragment to be inverted. The invention also relates to a method for the stable inversion of the DNA fragment upon rearrangement mediated by a recombinase such as Cre recombinase. The invention also relates to a method for obtaining a transgenic cell of which at least one allele of a DNA sequence of interest is invalidated by a process of conditional deletion and the genome of which has a reporter gene inserted at the place of the DNA fragment deleted by the process of conditional deletion. A method to generate targeting sites to perform site-specific recombination mediated cassette exchange is also provided.

REFERENCES:
patent: WO 00/63410 (2000-10-01), None
patent: WO 01/05987 (2001-01-01), None
patent: WO 02/40685 (2002-05-01), None
Berendsen, Hermann, Science. Oct. 1998, vol. 282, pp. 642-643.
Feng et al., “Site-specific Chromosomal Integration in Mammalian Cells: Highly Efficient CRE Recombinase-mediated Cassette Exchange,”J. Mol. Biol., 1999, pp. 779-785, vol. 292, No. 4, ©Academic Press, London, Great Britain.
Snaith et al., “Multiple cloning sites carryingloxPandFRTrecognition sites for the Cre and Flp site-specific recombinases,”Gene, 1995, pp. 173-174, vol. 166, No. 1, © Elsevier Science B.V., Amsterdam, Netherlands.
Wild et al., “Targeting and retrofitting pre-existing libraries of transposon insertions withFRTandoriVelements for in-vivo generation of large quantities of any genomic fragment,”Gene, 1998, pp. 55-66, vol. 223, No. 1-2, © Elsevier Science B.V., Amsterdamn, Netherlands.

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