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Last Updated: March 28, 2024

Claims for Patent: 5,814,716


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Summary for Patent: 5,814,716
Title: Cell lines from a transgenic mouse which express biologically active IX factor
Abstract:Cell lines have been prepared from transgenic mice comprised of a first exogenous DNA sequence encoding human factor IX which is controlled by a promoter region specific for hepatic cells and a second exogenous DNA sequence encoding an oncogene, such as the SV40 virus gene encoding the T-antigen or the mouse c-myc gene, which is controlled by a promoter region specific for hepatic cells. Promoter regions specific for hepatic cells included the promoter region for the factor IX gene and the promoter region for the .alpha.-1 antitrypsin gene. The cultured cells produce biologically active human factor IX which is then isolated from the culture media.
Inventor(s): Jallat; Sophie (Strasbourg, FR), Meulien; Pierre (Paris, FR), Pavirani; Andrea (Strasbourg, FR), Perraud; Frederic (Strasbourg, FR)
Assignee: Transgene S.A. (Courbevoie, FR)
Application Number:08/394,210
Patent Claims:1. A transgenic mouse which develops a liver tumor, wherein the tumor produces biologically active human factor IX, said mouse having integrated into its genome:

(i) a first exogenous DNA sequence encoding human factor IX, wherein said DNA sequence is genomic DNA, and where expression of said DNA sequence is controlled by a promoter region specific for hepatic cells, wherein said promoter region is the factor IX gene promoter or a portion thereof sufficient to provide for expression of said DNA sequence; and

(ii) a second exogenous DNA sequence encoding an oncogene, wherein said oncogene is selected from the group consisting of the SV40 virus gene encoding T-antigen and the mouse c-myc gene, where expression of the said DNA sequence is controlled by a promoter region specific for hepatic cells,

wherein said promoter region is an .alpha.-1 antitrypsin gene promoter,

wherein a cell line comprising TMhepTG48 is established from said mouse.

2. A cell line established from hepatic tissue of a transgenic mouse according to claim 1, wherein said cell line expresses biologically active human factor IX, said cell line having integrated in its genome:

(i) a first exogenous DNA sequence encoding human factor IX, wherein said DNA sequence is genomic DNA, and where expression of said DNA sequence is controlled by a promoter region specific for hepatic cells, wherein said promoter region is the factor IX gene promoter or a portion thereof sufficient to provide for expression of said DNA sequence; and

(ii) a second exogenous DNA sequence encoding an oncogene, wherein said oncogene is selected from the group consisting of the SV40 virus gene encoding T-antigen and the mouse c-myc gene, where expression of the said DNA sequence is controlled by a promoter region specific for hepatic cells, and wherein said promoter region is an .alpha.-1 antitrypsin gene promoter,

wherein said cell line comprises TMhepTG48.

3. A method for preparing a biologically active human factor IX, in which a cell line according to claim 2 is cultured and said factor IX is harvested from the culture medium.

4. A cell line established from hepatic tissue of a transgenic mouse according to claim 2, wherein said cell line expresses biologically active human factor IX, said cell line having integrated into its genome:

(i) a first exogenous DNA sequence encoding human factor IX, wherein said DNA sequence is genomic DNA comprising at least one intron, and where expression of said DNA sequence is controlled by a promoter region specific for hepatic cells, wherein said promoter region is the factor IX gene promoter or a portion thereof sufficient to provide for expression of said DNA sequence; and

(ii) a second exogenous DNA sequence encoding an oncogene, wherein said oncogene is selected from the SV40 virus gene encoding T-antigen and the mouse c-myc gene, where expression of the said DNA sequence is controlled by a promoter region specific for hepatic cells, wherein said promoter region is an .alpha.-l antitrypsin gene promoter,

wherein said cell line comprises TMhepTG48.

5. A cell line according to claim 2, wherein said first exogenous DNA sequence is a minigene.

6. A transgenic mouse according to claim 1, wherein said first exogenous DNA sequence is a minigene.

7. A method for preparing a cell line comprising removing the tumorous liver of a transgenic mouse according to claim 1, and culturing the cells of said liver to select a cell line, wherein said cell line produces biologically active human factor IX, said cell line having integrated into its genome:

(i) a first exogenous DNA sequence encoding human factor IX, wherein said DNA sequence is genomic DNA comprising at least one intron, and where expression of said DNA sequence is controlled by a promoter region specific for hepatic cells, wherein said promoter region is the factor IX gene promoter or a portion thereof sufficient to provide for expression of said DNA sequence; and

(ii) a second exogenous DNA sequence encoding an oncogene, wherein said oncogene is selected from the group consisting of the SV40 virus gene encoding T-antigen and the mouse c-myc gene, where expression of the said DNA sequence is controlled by a promoter region specific for hepatic cells, wherein said promoter region is an .alpha.-1 antitrypsin gene promoter,

wherein said cell line comprises TMhepTG48.

8. A transgenic mouse which has integrated into its genome:

(i) a first exogenous DNA sequence encoding human factor IX, wherein said DNA sequence is genomic DNA comprising at least one intron, and where expression of said DNA sequence is controlled by a promoter region specific for hepatic cells;

and

(ii) a second exogenous DNA sequence encoding an oncogene, wherein said oncogene is selected from the group consisting of the SV40 virus gene encoding T-antigen and the mouse c-myc gene, where expression of the said DNA sequence is controlled by a promoter region specific for hepatic cells,

wherein said mouse produces biologically active human factor IX.

9. A transgenic mouse of claim 8, wherein said first exogenous DNA sequence is a minigene.

10. A cell line established from hepatic tissue of a transgenic mouse according to claim 8, wherein said cell line expresses biologically active human factor IX.

11. A cell line according to claim 10, wherein said first exogenous DNA sequence is a minigene.

12. A method for preparing a biologically active human factor IX, in which a cell line according to claim 10 is cultured and said factor IX is harvested from the culture medium.

Details for Patent 5,814,716

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 06/04/1986 ⤷  Try a Trial 2015-09-29
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 ⤷  Try a Trial 2015-09-29
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b Injection 103132 ⤷  Try a Trial 2015-09-29
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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