Production of antibodies using cre-mediated site-specific recombination
First Claim
1. An embryonic stem cell of a mouse comprising a genome comprising:
- at least a functional portion of a genomically integrated human heavy chain immunoglobulin locus or at least a functional portion of a genomically integrated human light chain immunoglobulin locus,wherein a lox site is 5′
of said genomically integrated human heavy chain or said genomically integrate light chain immunoglobulin locus.
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Accused Products
Abstract
A method to produce a cell expressing an antibody from a genomic sequence of the cell comprising a modified immunoglobulin locus using Cre-mediated site-specific recombination is disclosed. The method involves first transfecting an antibody-producing cell with a homology-targeting vector comprising a lox site and a targeting sequence homologous to a first DNA sequence adjacent to the region of the immunoglobulin loci of the genomic sequence which is to be converted to a modified region, so the first lox site is inserted into the genomic sequence via site-specific homologous recombination. Then the cell is transfected with a lox-targeting vector comprising a second lox site suitable for Cre-mediated recombination with the integrated lox site and a modifying sequence to convert the region of the immunoglobulin loci to the modified region. This conversion is performed by interacting the lox sites with Cre in vivo, so that the modifying sequence inserts into the genomic sequence via Cre-mediated site-specific recombination of the lox sites. Also disclosed are a form of the method used to produce a cell expressing a modified antibody molecule using Cre-mediated site-specific recombination, and antibody-producing cells obtainable by the disclosed methods. Class-switching modifications of human antibodies produced in murine hybridoma cells are exemplified.
55 Citations
2 Claims
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1. An embryonic stem cell of a mouse comprising a genome comprising:
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at least a functional portion of a genomically integrated human heavy chain immunoglobulin locus or at least a functional portion of a genomically integrated human light chain immunoglobulin locus, wherein a lox site is 5′
of said genomically integrated human heavy chain or said genomically integrate light chain immunoglobulin locus.
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2. A transgenic mouse comprising a genome comprising:
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at least a functional portion of a genomically integrated human heavy chain immunoglobulin locus or at least a functional portion of a genomically integrated human light chain immuno globulin locus, wherein a lox site is 5′
of said genomically integrated human heavy chain or said genomically integrated light chain immunoglobulin locus.
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Specification