US 2003/0159160 A1 · Method for the stable inversion of DNA sequence by site-specific recombination and DNA vectors and transgenic cells thereof

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 said DNA fragment to be inverted. The invention also relates to a method for the stable inversion of said 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 comprises a reporter gene inserted at the place of the DNA fragment deleted by said process of conditional deletion. The invention also concerns a method to generate targeting sites to perform site-specific recombination mediated cassette exchange. The corresponding vectors, host cells, and transgenic animals are claimed.

Claims52

Dependent claims

An isolated DNA molecule comprising at least a sequence A flanked by at least site specific recombinase targeting sequences (SSRTS) L1, and at least a sequence B flanked by at least site specific recombinase targeting sequences (SSRTS) L2, said SSRTS L1 and SSRTS L2 being unable to recombine with one another, and wherein:

(i) sequences L1 are in an opposite orientation, and
(ii) sequences L2 are in an opposite orientation, and
(iii) the order of SSRTS sequences in said DNA molecule is 5′-L1-L2-L1-L2-3′.

Dependencies are inferred from the claim text.

Facts

Application

09/843,150Filed 27 Apr 2001

Published

21 Aug 2003

Term ends

Not estimatedThe source records don’t support an estimate.

Classification8 codes

C12N 15/90
Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation: stable introduction of foreign DNA…
C12N 15/63
Recombinant DNA-technology: introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
C12N 2800/30
Nucleic acids vectors: vector systems comprising sequences for excision in presence of a recombinase, e.g. loxP or FRT
C12N 2800/60
Nucleic acids vectors: vectors containing traps for, e.g. exons, promoters

Method for the stable inversion of DNA sequence by site-specific recombination and DNA vectors and transgenic cells thereof

Life of this patent

Method for the stable inversion of DNA sequence by site-specific recombination and DNA vectors and transgenic cells thereof

Life of this patent

Abstract

Claims52