Patent ID: 6458592
Filing Date: 2002-10-01
Classification: A01K,C07K,C12N

Abstract:
A method of producing an antibody-producing cell, said cell comprising a genomically-integrated modified immunoglobulin locus, which method comprises the steps of:(a) transfecting an antibody-producing cell in vitro with a first homology-targeting vector comprising: (i) a first lox site, and (ii) a targeting sequence homologous to a first DNA sequence 5â€² of a region of a genomically-integrated immunoglobulin locus in said cell, so that said first lox site is inserted 5â€² of said region via homologous recombination with the genomically-integrated immunoglobulin locus, (b) transfecting said cell of step (a) in vitro with a lox-targeting vector comprising: (i) a second lox site suitable for Cre-mediated recombination with said first lox site, and (ii) a modifying sequence; (c) interacting the lox sites with Cre, so that said modifying sequence inserts into said immunoglobulin locus via Cre-mediated site-specific recombination of said lox sites, thereby forming a modified immunoglobulin locus; and (d) selecting a transfectant in vitro, which transfectant comprises said modified immunoglobulin locus and which transfectant produces a functional antibody molecule.