Methods for producing recombinant mammalian cells harboring a yeast artificial chromosome
AssigneeGenpharm Internation, Inc.
InventorsJeanne F. Loring, Theodore Choi, Robert M. Kay
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Abstract
The present invention provides methods and compositions for transferring large transgene polynucleotides and unlinked selectable marker polynucleotides into eukaryotic cells by a novel method designated co-lipofection. The methods and compositions of the invention are used to produce novel transgenic non-human animals harboring large transgenes, such as a transgene comprising a human APP gene or human immunoglobulin gene.
Claims11
A method for producing a selectable co-lipofected mammalian cell having incorporated multiple heterologous DNA species, said mammalian cell having a genome comprising the steps of:
- forming a co-lipofection complex comprising a cationic lipid, a first polynucleotide, said first polynucleotide being larger than 50 Kb, and an unlinked second polynucleotide comprising a selectable marker gene expression cassette;
- contacting a mammalian cell with said co-lipofection complex under conditions whereby said first polynucleotide and said second polynucleotide are both introduced into said cell and are integrated into said genome to form a selectable co-lipofected mammalian cell having incorporated multiple heterologous DNA species.
Dependencies are inferred from the claim text.