Methods for identifying human cell lines useful for endogenous gene activation, isolated human lines identified thereby, and uses thereof

An isolated, human recombinant cell produced by endogenous gene activation wherein said human recombinant cell comprises a heterologous promoter in operable linkage with said endogenous gene, wherein said endogenous gene encodes erythropoietin, wherein said CMV promoter and said endogenous gene are amplified and wherein 106 of said cells produce at least 1000 ng of erythropoietin per 24 hours.

An isolated, human recombinant cell which comprises a CMV promoter sequence in operable linkage with an endogenous gene which encodes a protein, wherein 106 of said cell produces at least 200 ng of said protein per 24 hours, wherein said endogenous gene is not amplified, wherein the isolated human recombinant cell is produced by endogenous gene activation of an endogenous gene in a human cell that comprises more than two chromosomes containing the endogenous gene and wherein the human cell does not express the endogenous gene prior to endogenous gene activation, wherein said human cell undergoes 5 doublings over a period of 14 days or less in suspension culture, 5 doublings over a maximum period of fourteen days in a serum free culture medium and is capable of correctly glycosylating the protein.

An isolated, human recombinant cell which comprises a CMV promoter sequence in operable linkage with an endogenous gene which encodes a protein, wherein 106 of said cell produces at least 200 ng of said protein per 24 hours, wherein said endogenous gene is not amplified, wherein the isolated human recombinant cell is produced by endogenous gene activation of an endogenous gene encoding erythropoietin (EPO) in a human cell that comprises more than two chromosomes containing the endogenous gene encoding EPO and wherein the human cell does not express the endogenous gene prior to endogenous gene activation.

An isolated, human recombinant immortalized cell which comprises a CMV promoter sequence in operable linkage with an endogenous gene which encodes a protein, wherein 106 of said cell produces at least 200 ng of said protein per 24 hours, wherein said endogenous gene is not amplified, wherein the isolated human recombinant immortalized cell is produced by endogenous gene activation of an endogenous gene in a human immortalized cell that comprises more than two chromosomes containing the endogenous gene and wherein the human cell does not express the endogenous gene prior to endogenous gene activation.