Production of antibodies using Cre-mediated site-specific recombination
US6130364A · kind A · utility
Assignees
Inventors
Key dates
| Filing date | Mar 29, 1995 |
| Grant date | Oct 10, 2000 |
| Priority date | — |
| Expiry date | Mar 29, 2015 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2800/30
- WIPO fieldBiotechnology
- WIPO sectorChemistry
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 recombinati…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.