Activating expression of an amplifying endogenous gene by...

Details Activating expression of an amplifying endogenous gene by... Activating expression of an amplifying endogenous gene by...

C - Chemistry, Metallurgy

12

N

C12N 15/90 (2006.01) A61K 35/12 (2006.01) A61K 48/00 (2006.01) C07K 14/505 (2006.01) C07K 14/61 (2006.01) C12N 1/19 (2006.01) C12N 1/21 (2006.01) C12N 5/10 (2006.01) C12N 11/00 (2006.01) C12N 15/11 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2151032

Method of activating expression of and amplifying an endogenous gene in genomic DNA of a vertebrate cell which is not expressed in the cell as obtained or is not expressed at significant levels in the cell as obtained, comprising the steps of: a) transfecting cells with DNA sequences comprising exogenous DNA sequence which repair, alter, delete or replace a sequence present in the cell or which are regulatory sequences not normally functionally linked to the endogenous gene in the cell as obtained; 2) DNA sequences homologous with genomic DNA sequences at a preselected site in the cells; and 3) amplifiable DNA encoding a selectable marker, b) maintaining the cells under conditions appropriate for homologous recombination to occur between DNA sequences homologous with genomic DNA sequences and genomic DNA sequences, c) culturing the homologously recombinant cells produced in (b) under conditions which select for amplification of the amplifiable DNA encoding a selectable marker, whereby the amplifiable DNA encoding a selectable marker and the endogenous gene functionally linked to exogenous DNA are coamplified.

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