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Last Updated: April 25, 2024

Claims for Patent: 5,565,362

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Summary for Patent: 5,565,362

Title:	DNA sequences to target proteins to the mammary gland for efficient secretion
Abstract:	Described is a method of targeting specific genes to the mammary gland which results in the efficient synthesis and secretion of biologically important molecules. Further, there is described as a composition of matter, a transgenic mammal having the ability to reproduce itself and being suitable for the secretion of biologically active agents into its milk. Additionally there is disclosed as a composition of matter, recombinant DNA gene complexes designed to integrate into a mammalian genome and to synthesize and secrete biological active agents into the milk. Furthermore methods of producing and using altered milk are disclosed.
Inventor(s):	Rosen; Jeffrey M. (Houston, TX)
Assignee:	Pharming B.V. (Leiden, NL)
Application Number:	08/185,574
Patent Claims:	1. A recombinant DNA complex to provide preferential mammary gland expression and secretion of a selected polypeptide comprising: (a) a casein promoter; (b) a transcribable DNA segment coding for said selected polypeptide, wherein said promoter does not naturally control the transcription of said DNA segment; (c) secretory DNA encoding a signal peptide functional in mammary secretory cells operably linked between said promoter and said DNA segment to facilitate secretion of said selected polypeptide; (d) a casein enhancer sequence capable of conferring enhanced preferential transcription of said recombinant DNA complex in mammary secretory cells when said complex is integrated into the genome of a non-human mammal, and (e) an intronic sequence from a casein gene or said transcribable DNA segment. 2. The recombinant DNA complex of claim 1 wherein said promoter is from a casein gene selected from the group consisting of .alpha.-casein, .beta.-casein, .gamma.-casein and .kappa.-casein. 3. The recombinant DNA complex of claim 2 wherein said promoter comprises the promoter from a .beta.-casein gene. 4. The recombinant DNA complex of claim 3 wherein said casein enhancer is selected from the group consisting of .alpha.-casein, .beta.-casein, .gamma.-casein and .kappa.-casein enhancers. 5. The recombinant DNA complex of claim 1 further comprising a mouse mammary tumor virus glucocorticoid response element. 6. The recombinant DNA complex of claim 1 wherein said secretory DNA encodes a signal peptide from a mammary gland specific gene. 7. The recombinant DNA complex of claim 6 wherein said mammary gland specific gene is selected from the group consisting of whey acid protein and casein genes. 8. The recombinant DNA complex of claim 7 wherein said mammary gland specific gene is a casein gene. 9. The recombinant DNA complex of claim 8 wherein said casein gene is selected from the group consisting of .alpha.-casein, .beta.-casein, and .kappa.-casein genes. 10. The recombinant DNA complex of claim 1 wherein said transcribable DNA segment encodes a normally secreted polypeptide and said secretory DNA encodes the signal sequence of said secreted polypeptide. 11. The recombinant DNA complex of claim 1 further comprising: (f) DNA encoding a 5' untranslated sequence from a mammary gland specific gene. 12. The recombinant DNA complex of claim 11 further comprising: (g) DNA encoding a 3' untranslated sequence. 13. The recombinant DNA complex of claim 1, wherein said intronic sequence links said transcribable DNA segment to said promoter. 14. The recombinant DNA complex of claim 1, wherein said intronic sequence is located within said transcribable DNA segment. 15. The recombinant DNA complex of claim 13, wherein said intronic sequence is a portion of an intron located between exons I and II of a rat .beta.-casein gene. 16. A vector to provide preferential mammary gland expression and secretion of a selected polypeptide comprising: (a) a casein promoter; (b) a transcribable DNA segment coding for said selected polypeptide, wherein said promoter does not naturally control the transcription of said DNA segment; and (c) secretory DNA encoding a signal peptide functional in mammary secretory cells operably linked between said promoter and said DNA segment to facilitate secretion of said selected polypeptide, (d) a casein enhancer sequence capable of conferring enhanced preferential transcription of said recombinant DNA complex in mammary secretory cells when said complex is integrated into the genome of a non-human mammal, and (e) an intron from a casein gene. 17. A recombinant DNA complex to provide preferential mammary gland expression and secretion of a selected polypeptide comprising: (a) a casein promoter; (b) a transcribable DNA segment coding for said selected polypeptide, wherein said promoter does not naturally control the transcription of said DNA segment; and (c) secretory DNA encoding a signal peptide functional in mammary secretory cells operably linked between said promoter and said DNA segment to facilitate secretion of said selected polypeptide, (d) a casein enhancer sequence capable of conferring enhanced preferential transcription of said recombinant DNA complex in mammary secretory cells when said complex is integrated into the genome of a non-human mammal, (e) an intron from a casein gene; and (f) a restriction enzyme linker sequence. 18. A recombinant DNA complex to provide preferential mammary gland expression and secretion of a selected polypeptide comprising: (a) a casein promoter; (b) a transcribable DNA segment coding for said selected polypeptide, wherein said promoter does not naturally control the transcription of said DNA segment; and (c) secretory DNA encoding a signal peptide functional in mammary secretory cells operably linked between said promoter and said DNA segment to facilitate secretion of said selected polypeptide, (d) a casein enhancer sequence capable of conferring enhanced preferential transcription of said recombinant DNA complex in mammary secretory cells when said complex is integrated into the genome of a non-human mammal, and (e) an intron from a casein gene; wherein the recombinant DNA complex is linear.

Details for Patent 5,565,362

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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	2013-10-15
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132		⤷ Try a Trial	2013-10-15
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132		⤷ Try a Trial	2013-10-15
>Applicant	>Tradename	>Biologic Ingredient	>Dosage Form	>BLA	>Approval Date	>Patent No.	>Expiredate

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