Claims for Patent: 6,200,778

Summary for Patent: 6,200,778

Title:	Genomic sequences for protein production and delivery
Abstract:	An isolated nucleic acid molecule that hybridizes under stringent conditions, or shares at least 80% sequence identity, with a defined genomic region upstream of the coding region of a FSH.beta. gene, and a DNA construct containing that nucleic acid molecule as a targeting sequence for homologous recombination.
Inventor(s):	Treco; Douglas A. (Arlington, MA), Heartlein; Michael W. (Boxborough, MA), Selden; Richard F (Wellesley, MA)
Assignee:	Transkaryotic Therapies, Inc. (Cambridge, MA)
Application Number:	09/305,639
Patent Claims:	1. A DNA construct that alters expression of an endogenous FSH.beta. gene in a mammalian cell upon integration into the genome of the cell via homologous recombination, the construct comprising a targeting sequence containing at least 20 contiguous nucleotides from SEQ ID NO:4 and at least 20 contiguous nucleotides from SEQ ID NO:5, and a transcriptional regulatory sequence. 2. The DNA construct of claim 1, wherein the construct further comprises an exon and a splice-donor site. 3. The DNA construct of claim 2, wherein the construct further comprises, downstream from the splice-donor site, an intron and a splice-acceptor site. 4. A homologously recombinant cell stably transfected with the DNA construct of claim 3, the DNA construct having undergone homologous recombination with genomic DNA upstream of the ATG initiation codon of an endogenous FSH.beta. coding sequence. 5. A method of producing FSH.beta., comprising providing the cell of claim 4, and culturing the cell in vitro under conditions which permit the cell to express and secrete FSH.beta.. 6. A homologously recombinant cell stably transfected with the DNA construct of claim 2, the DNA construct having undergone homologous recombination with genomic DNA upstream of the ATG initiation codon of an endogenous FSH.beta. coding sequence. 7. A method of producing FSH.beta. comprising providing the cell of claim 6, and culturing the cell in vitro under conditions which permit the cell to express and secrete FSH.beta.. 8. The DNA construct of claim 1, wherein the construct further comprises a selectable marker gene. 9. A homologously recombinant cell stably transfected with the DNA construct of claim 8, the DNA construct having undergone homologous recombination with genomic DNA upstream of the ATG initiation codon of an endogenous FSH.beta. coding sequence. 10. A method of producing FSH.beta., comprising providing the cell of claim 9, and culturing the cell in vitro under conditions which permit the cell to express and secrete FSH.beta.. 11. The DNA construct of claim 1, wherein the targeting sequence contains at least 50 contiguous nucleotides from SEQ ID NO:4 or 5. 12. A homologously recombinant cell stably transfected with the DNA construct of claim 1, the DNA construct having undergone homologous recombination with genomic DNA upstream of the ATG initiation codon of an endogenous FSH.beta. coding sequence. 13. A method of producing FSH.beta., comprising providing the cell of claim 12, and culturing the cell in vitro under conditions which permit the cell to express and secrete FSH.beta.. 14. A method of altering expression of an endogenous FSH.beta. gene in a mammalian cell in vitro, the method comprising introducing the DNA construct of claim 1 into the cell; maintaining the cell under conditions which permit homologous recombination to occur between the construct and a genornic target site homologous to the targeting sequence, to produce a homologously recombinant cell; and maintaining the homologously recombinant cell under conditions which permit expression of the FSH.beta. coding sequence under the control of the transcriptional regulatory sequence. 15. An isolated nucleic acid comprising at least 100 contiguous nucleotides of SEQ ID NO:4 or its complement. 16. The isolated nucleic acid of claim 15, wherein the isolated nucleic acid comprises at least 200 contiguous nucleotides of SEQ ID NO:4 or its complement. 17. The isolated nucleic acid of claim 15, wherein the isolated nucleic acid comprises at least 500 contiguous nucleotides of SEQ ID NO:4 or its complement. 18. The isolated nucleic acid of claim 15, wherein the isolated nucleic acid comprises at least 1000 contiguous nucleotides of SEQ ID NO:4 or its complement. 19. The isolated nucleic acid of claim 15, wherein the isolated nucleic acid comprises SEQ ID NO:4 or its complement. 20. An isolated nucleic acid comprising at least 50 contiguous nucleotides of SEQ ID NO:5 or its complement. 21. The isolated nucleic acid of claim 20, wherein the isolated nucleic acid comprises at least 100 contiguous nucleotides of SEQ ID NO:5 or its complement. 22. The isolated nucleic acid of claim 20, wherein the isolated nucleic acid comprises at least 200 contiguous nucleotides of SEQ ID NO:5 or its complement. 23. The isolated nucleic acid of claim 20, wherein the isolated nucleic acid comprises SEQ ID NO:5 or its complement. 24. A DNA construct that alters expression of an endogenous FSH.beta. gene in a mammalian cell upon integration into the genome of the cell via homologous recombination, the construct comprising a targeting sequence containing at least 20 contiguous nucleotides from SEQ ID NO:5 and a transcriptional regulatory sequence. 25. The DNA construct of claim 24, wherein the construct further comprises a exon and a splice-donor site. 26. The DNA construct of claim 25, wherein the construct further comprises, downstream from the splice-donor site, an intron and a splice-acceptor site. 27. A homologously recombinant cell stably tsfected with the DNA construct of claim 26, the DNA construct having undergone homologous recombination with genomic DNA upstream of the ATG initiation codon of an endogenous FSH.beta. coding sequence. 28. A method of producing FSH.beta., comprising providing the cell of claim 27, and culturing the cell in vitro under conditions which permit the cell to express and secrete FSH.beta.. 29. A homologously recombinant cell stably transfected with the DNA construct of claim 25, the DNA construct having undergone homologous recombination with genomic DNA upstream of the ATG initiation codon of an endogenous FSH.beta. coding sequence. 30. A method of producing FSH.beta., comprising providing the cell of claim 29, and culturing the cell in vitro under conditions which permit the cell to express and secrete FSH.beta.. 31. The DNA construct of claim 24, wherein the construct further comprises a selectable marker gene. 32. A homologously recombinant cell stably transfected with the DNA construct of claim 31, the DNA construct having undergone homologous recombination with genomic DNA upstream of the ATG initiation codon of an endogenous FSH.beta. coding sequence. 33. A method of producing FSH.beta., comprising providing the cell of claim 32, and culturing the cell in vitro under conditions which permit the cell to express and secrete FSH.beta.. 34. The DNA construct of claim 24, wherein the targeting sequence contains at least 50 contiguous nucleotides from SEQ ID NO:5. 35. A homologously recombinant cell stably transfected with the DNA construct of claim 24, the DNA construct having undergone homologous recombination with genomic DNA upstream of the ATG initiation codon of an endogenous FSH.beta. coding sequence. 36. A method of producing FSH.beta., comprising providing the cell of claim 35, and culturing the cell in vitro under conditions which permit the cell to express and secrete FSH.beta.. 37. A method of altering expression of an endogenous FSH.beta. gene in a mammalian cell in vitro, the method comprising, introducing the DNA construct of claim 24, into the cell; maintaining the cell under conditions which permit homologous recombination to occur between the construct and a genomic target site homologous to the targeting sequence, to produce a homologously recombinant cell; and maintaining the homologously recombinant cell under conditions which permit expression of the FSH.beta. coding sequence under the control of the transcriptional regulatory sequence.

Details for Patent 6,200,778

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		2018-05-07
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2018-05-07
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2018-05-07