Patent ID: 6632672
Filing Date: 2003-10-14
Classification: A61K,C12N

Abstract:
A method of site-specifically integrating a polynucleotide sequence of interest in the genome of an isolated eucaryotic cell, said method comprising:introducing (i) a circular targeting construct, comprising a first recombination site and the polynucleotide sequence of interest, and (ii) an expression cassette comprising a polynucleotide encoding a site-specific recombinase into the isolated eucaryotic cell, wherein (a) the genome of said isolated eucaryotic cell comprises a second recombination site native to the genome, (b) recombination between the first and second recombination sites occurs in the presence of the site-specific recombinase, and (c) the site-specific recombinase is selected from the group consisting of &phgr;C31 phage recombinase, TP901-1 phage recombinase, and R4 phage recombinase; and maintaining the isolated eucaryotic cell under conditions that allow recombination between said first and second recombination sites, wherein the recombination is mediated by the site-specific recombinase and the recombination results in site-specific integration of the polynucleotide sequence of interest in the genome of the isolated eucaryotic cell.