Claims
- 1. An expression vector comprising a promoter operably linked to a βARK (Beta-Adrenergic Receptor Kinase) gene employed to effect overexpression of βARK.
- 2. An expression vector according to claim 1 wherein the promoter is the α-myosin heavy chain promoter, CMV promoter, β-myosin heavy chain promoter, or any other promoter capable of inducing said overexpression in myocardial cells.
- 3. An expression vector according to claim 2 wherein the βARK gene is a βARK1 gene.
- 4. The expression vector according to claim 3 wherein the βARK1 gene is bovine βARK1.
- 5. An expression vector according to claim 1 further characterized as containing a stabilizing Simian virus 40 (SV-40) sequence at the 3′ end of the βARK gene.
- 6. An expression vector for mouse myocardial cell expression comprising the murine alpha-myosin heavy chain (α-MHC) promoter operably linked to a DNA sequence encoding bovine βARK1 and terminating with a portion of the SV-40 intron.
- 7. An expression vector according to claim 1 wherein the vector further comprises a replication deficient recombinant adenovirus.
- 8. A composition suitable for use in gene therapy comprising the expression vector of any one of claims 1 through 7 contained in a physiologically acceptable carrier.
- 9. A myocardial cell transfected by the expression vector of any of claims 1-7.
- 10. A transgenic mouse, whose germ cells and somatic cells contain a beta adrenergic receptor kinase gene which is operably linked to a promoter effective for the expression of said gene in the myocardial tissue of said mouse and effective for increasing the amount of β-adrenergic receptor kinase in myocardium of said mouse, said gene being introduced into said mouse, or an ancestor of said mouse containing said gene.
- 11. The mouse of claim 10, wherein said promoter sequence controlling expression of said gene is the murine alpha myosin heavy chain promoter.
- 12. The mouse of claim 10, wherein said expressed gene is operably linked to a SV 40 stabilizing sequence at the 3′ end.
- 13. A method of assessing the effectiveness of a compound in augmenting cardiac function comprising
administering to an adult transgenic animal according to any one of claims 10-12 an amount of the compound, measuring a biochemical or physiological parameter indicative of effectiveness, and assessing the effectiveness by comparing the measured parameter value to a base line reading.
- 14. An expression vector comprising a promoter operably linked to a DNA sequence encoding a βARK (Beta-Adrenergic Receptor kinase) inhibitor.
- 15. An expression vector according to claim 14 wherein the promoter is the α-myosin heavy chain promoter, CMV promoter, β-myosin heavy chain promoter, or any other promoter capable of inducing expression in myocardial cells.
- 16. An expression vector according to claim 14 wherein the DNA sequence encodes the carboxyl terminus of βARK1.
- 17. An expression vector according to claim 16 wherein the DNA sequence encodes about 200 amino acids at the carboxyl terminus of βARK1.
- 18. An expression vector according to claim 14 further characterized as containing a stabilizing Simian virus 40 (SV-40) sequence at the 3′ end of the DNA sequence.
- 19. An expression vector for mouse myocardial cell expression comprising the murine alpha-myosin heavy chain (α-MHC) promoter operably linked to a DNA sequence encoding the carboxyl terminus of βARK1 and terminating with a portion of the SV-40 intron.
- 20. An expression vector according to claim 14 wherein the vector further comprises a replication deficient recombinant adenovirus.
- 21. A composition suitable for use in gene therapy comprising the expression vector construct of any one of claims 14 through 20 contained in a physiologically acceptable carrier and in an effective amount to augment cardiac function.
- 22. A myocardial cell transfected by the expression vector of any of claims 14-20.
- 23. A transgenic mouse, whose germ cells and somatic cells contain an exogenous DNA sequence encoding a beta adrenergic receptor kinase inhibitor which is operably linked to a promoter effective for the expression of said gene in the myocardial tissue of said mouse and effective for enhancing myocardial function in said mouse, said DNA being introduced into said mouse, or an ancestor of said mouse containing said gene.
- 24. The mouse of claim 23, wherein said promoter sequence controlling expression of said DNA sequence is the murine alpha myosin heavy chain promoter.
- 25. The mouse of claim 23, wherein said expressed DNA sequence is operably linked to a SV 40 stabilizing sequence at the 3′ end.
- 26. A method of improving myocardial function in mammals comprising administering to said mammals an effective amount of a transgene (exogenous gene or foreign DNA sequence) comprising
a myocardial promoter operably linked to a DNA sequence encoding a beta adrenergic receptor kinase inhibitor such that an increase in myocardial function is effected.
- 27. The method of improving myocardial function in mammals according to claim 24 wherein said DNA sequence comprises the carboxyl terminus of bovine βARK1, or an allelic variation thereof.
- 28. The method of improving myocardial function in mammals according to claim 24 wherein said transgene is a vector.
- 29. The method of improving heart function in mammals according to claim 24 wherein said myocardial promoter is selected from the group consisting of: the CMV promoter, Rous sarcoma virus promoter, β-myosin heavy chain promoter, and α-myosin heavy chain promoter.
- 30. A method for improving heart function in a mammal comprising administering to said mammal myocardial cells according to claim 20 such that the function of β-AR in said heart is augmented.
- 31. A mouse having a phenotype characterized by an elevated level of β2 adrenergic receptors in myocardium, and increased chronotropy and/or inotropy from that naturally occuring in said mouse.
- 32. The mouse of claim 31, said phenotype being conferred by a transgene contained in the somatic and germ cells of said mouse, said transgene comprising the human β2-AR gene operably linked to the α-myosin heavy chain promoter.
- 33. A mouse having a phenotype characterized by an elevated level of βARK in myocardium, and decreased chronotropy and/or inotropy from that naturally occuring in said mouse.
- 34. The mouse of claim 33, said phenotype being conferred by a transgene contained in the somatic and germ cells of said mouse, said transgene comprising a βARK1 gene operably linked to the α-myosin heavy chain promoter.
- 35. A mouse having a phenotype characterized by the presence of a βARK inhibitor in myocardium, and increased chronotropy and/or inotropy from that naturally occuring in said mouse.
- 36. The mouse of claim 35, said phenotype being conferred by a transgene contained in the somatic and germ cells of said mouse, said transgene comprising a DNA sequence encoding the last 200 amino acids of βARK1 operably linked to the α-myosin heavy chain promoter.
Parent Case Info
[0001] This patent application is a continuation-in-part of application Ser. No. 08/228,012, filed Apr. 15, 1994.
Government Interests
[0002] The work described in this patent application was supported in part by NIH grants HL-16037, HL 5F32-CA09350 and by the American Heart Association, California affiliate grant 94-286. The government has certain rights in the invention.
Continuations (2)
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10183344 |
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08453202 |
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Continuation in Parts (1)
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08228012 |
Apr 1994 |
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08453202 |
May 1995 |
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