REGULATION OF ERYTHROPOIETIN EXPRESSION BY DOXYCYCLINE IN RAT PRIMARY SKELETAL MUSCLE CELLS FOLLOWING TRANSDUCTION BY TETRACYCLINE GENE EXPRESSION SYSTEM

Dai Yong; Xu Zhuojia; Li Tiyuan

doi:10.29333/ejgm/82389

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Dai Yong ¹ ^* , Xu Zhuojia ¹, Li Tiyuan ¹

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¹ 2nd Affiliated Hospital of Medical College of Jinan University, Shenzhen People’s Hospital, China^* Corresponding Author

Abstract

Aim: Future prospects for gene therapy of renal anemia involve expression of the EPO transgene. Control of gene expression is important to gene therapy for purposes of both dosing and safety. We tested the effects of rhEPO gene expression under the tetracycline gene expression system’s regulating and controlling. Methods: Cultured rat primary skeletal muscle cells for gene delivery use; Inserted the hEPO cDNA in the plasmid pTRE2hyg; Rat primary skeletal muscle cells were engineered for Dox-inducible and skeletal muscle-specific expression of rhEPO cDNA by co-transfection pTRE2hyg-EPO and Tet-ON plasmids. Results: EPO could be secreted by the transfected positive cells. EPO expression depended on the presence of doxycycline, and the EPO secretion was increased parallel to dose of doxycycline. Conclusion: Tetracycline gene expression system provided a suitable control system for sustained and regulated expression of a desired gene.

Keywords

EPO
Dox
Gene therapy
Skeletal Muscle cells

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Article Type: Original Article

EUR J GEN MED, Volume 3, Issue 3, July 2006, 108-115

https://doi.org/10.29333/ejgm/82389

Publication date: 15 Jul 2006

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References

Koury MJ, Bondurant MC. The molecular mechanism of erythropoietin action. Eur J Biochem 1992;210:649-63
Goodnough LT, Skikne B, Brugnara C. Erythropoietin, iron, and erythropoiesis. Blood 2000;96:823-33
Krantz,S.B. Erythropoietin. Blood, 1991;77:419-34
Paganini, N.F.,& Miller, T. Erythropoietin therapy in renal failure. Adv Intern 1993;38:223-43
Domenico M, Maciej W, Paola D, et al. Prolonged expression and effective readministration of erythropoietin delivered with a fully deleted adenoviral vector. Hum Gene Ther 2000;11:859-68
Chenuaud P, Larcher T, Rabinowitz JE, et al. Autoimmune anemia in macaques following erythropoietin gene therapy. Blood, 2004;103:3303-4
Bohl D, Heard JM. Delivery erythropoietin through genetically engineered cells. J Am Soc Nephrol 2000;16:159-62
Schrier S. Pathobiology of thalassemic erythrocytes. Current Op Hematol 1997;4:75-8
Boh D, Bosch A, et. Improvement of erythropoiesis in beta-thalassemic mice by continuous erythropoietin delivery from muscle. Blood 2000;95:2793-8
Rizzuto G,Cappelletti M, et al. Gene electrotransfer results in a high-level teansduction of rat skeletal muscle and corrects anemia of renal failure. Hum Gene Ther 2000;11:1891-900
Sevensson EC, Black HB,Dugger DL,et al. Long-term erythropoietin expression in rodents and non-human primates following intramuscular injection of a replication-defective adenoviral vector. Human Gene Therapy 1997;8:797-806
Zhou S, Murphy JE,et al. Adenoassociated virus-mediated delivery of erythropoietin leads to sustained elevation of hematocrit in nonhuman primates. Gene Ther 1998;5:665-70
Barr E. & Leiden J. M. Systemic delivery of recombinant proteins by genetically modified myoblasts. Science 1991;254: 1507-19
Dhawan J., Pan L. C., Pavlath G. K., Travis, M. A., Lanctot, A. M. & Blau, H. M. Systemic delivery of human growth hormone by injection of genetically engineered myoblasts. Science 1991; 254:1509-12
Osborne W. R., Ramesh N., Lau S., Clowes M. M., Dale D. C. & Clowes A. W. Gene therapy for long-term expression of erythropoietin in rats. Proc Natl Acad Sci USA 1995;92:8055-8
Chowdhury JR, Grossman M, Gupta S, et al. Long-term improvement of hypercholesterolemia after ex vivo gene therapy in LDLR-deficient rabbits. Science 1991;254:1802-5
Blau HM. & Springer ML. Musclemediated gene therapy. N Engl J Med 1995;333:1554-6
Yao S N & Kurachi K. Expression of human factor IX in mice after injection of genetically modified myoblasts. Proc Natl Acad Sci USA 1992;89:3357-61
Tripathy SK, Goldwasser E, Lu MM, Barr E. & Leiden JM. Stable delivery of physiologic levels of recombinant erythropoietin to the systemic circulation by intramuscular injection of replicationdefective adenovirus. Proc Natl Acad Sci. USA 1994;91:11557-61
Naffakh N, Pinset C, Montarras D, Li Z, Danos O. & Heard JM. Long-term secretion of therapeutic proteins from genetically modified skeletal muscles. Gene Ther 1996;7:11-21
Leiden JM. Gene therapy-promise, pitfalls, and prognosis. N Engl J Med 1995;333:871-3
Furth PA. Temporal control of gene expression in transgeneic mice by a tetracycline-responsive promotor. Proc Natl Acad Sci USA 1994;91:9302-6
Shockett P, Difilippantonio M, Hellman N& Schatz D. A modified tetracyclineregulated system provides autoregulatory, inducible gene expression in cultured cells and transgenic mice. Proc Natl Acad Sci USA 1995;92:6522-6
Passman, R.S.& Fishman, G.I. Regulated expression of gene expression in vivo after germline transfer. J Clin Invest 1994;94:2421-5
Schultze N, Burki, Y, Certa, U.& Bluethmann, H. Efficient control of gene expression by single integration of the tetracycline system in transgenic mice. Nature Biotech 1996;14:499-505
Vogt K, Bhabhra R, Rhodes JC, Askew DS. Doxycycline-regulated gene expression in the opportunistic fungal pathogen Aspergillus fumigatus. BMC Microbiology 2005;5:1471-80
Clackson T. Regulated gene expression systems. Gene Ther 2000;7:120–5
Toniatti C. Bujard H. Cortese R. Ciliberto G. Gene therapy progress and prospects: transcription regulatory systems. Gene Ther 2004;11:649–57
Gossen M. Bujard H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl Acad Sci USA 1992;89:5547–51
Agha-Mohammadi S. Lotze MT. Regulatable systems: applications in gene therapy and replicating viruses. J Clin Invest 2000;105:1177–83
Corbel SY. Rossi FM. Latest developments and in vivo use of the Tet system: ex vivo and in vivo delivery of tetracycline- regulated genes. Curr Opin Biotechnol 2002;13:448–52
Gossen M. Freundlieb S. Bender G. Muller G. Hillen W. Bujard H. Transcriptional activation by tetracyclines in mammalian cells. Science 1995;268:1766–69
Baron U. Gossen M. Bujard H. Tetracycline-controlled transcription in eukaryotes: novel transactivators with graded transactivation potential. Nucleic Acids Res 1997;25:2723–9
Urlinger S. Baron U. Thellmann M. Hasan M.T. Bujard H. Hillen W. Exploring the sequence space for tetracyclinedependent transcriptional activators: novel mutations yield expanded range and sensitivity. Proc Natl Acad Sci USA 2000;97:7963–8
Lutz R. Bujard H. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res 1997;25: 1203–10

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Vancouver

Yong D, Zhuojia X, Tiyuan L. REGULATION OF ERYTHROPOIETIN EXPRESSION BY DOXYCYCLINE IN RAT PRIMARY SKELETAL MUSCLE CELLS FOLLOWING TRANSDUCTION BY TETRACYCLINE GENE EXPRESSION SYSTEM. EUR J GEN MED. 2006;3(3):108-15. https://doi.org/10.29333/ejgm/82389

APA

Yong, D., Zhuojia, X., & Tiyuan, L. (2006). REGULATION OF ERYTHROPOIETIN EXPRESSION BY DOXYCYCLINE IN RAT PRIMARY SKELETAL MUSCLE CELLS FOLLOWING TRANSDUCTION BY TETRACYCLINE GENE EXPRESSION SYSTEM. European Journal of General Medicine, 3(3), 108-115. https://doi.org/10.29333/ejgm/82389

AMA

Yong D, Zhuojia X, Tiyuan L. REGULATION OF ERYTHROPOIETIN EXPRESSION BY DOXYCYCLINE IN RAT PRIMARY SKELETAL MUSCLE CELLS FOLLOWING TRANSDUCTION BY TETRACYCLINE GENE EXPRESSION SYSTEM. EUR J GEN MED. 2006;3(3), 108-115. https://doi.org/10.29333/ejgm/82389

Chicago

Yong, Dai, Xu Zhuojia, and Li Tiyuan. "REGULATION OF ERYTHROPOIETIN EXPRESSION BY DOXYCYCLINE IN RAT PRIMARY SKELETAL MUSCLE CELLS FOLLOWING TRANSDUCTION BY TETRACYCLINE GENE EXPRESSION SYSTEM". European Journal of General Medicine 2006 3 no. 3 (2006): 108-115. https://doi.org/10.29333/ejgm/82389

Harvard

Yong, D., Zhuojia, X., and Tiyuan, L. (2006). REGULATION OF ERYTHROPOIETIN EXPRESSION BY DOXYCYCLINE IN RAT PRIMARY SKELETAL MUSCLE CELLS FOLLOWING TRANSDUCTION BY TETRACYCLINE GENE EXPRESSION SYSTEM. European Journal of General Medicine, 3(3), pp. 108-115. https://doi.org/10.29333/ejgm/82389

MLA

Yong, Dai et al. "REGULATION OF ERYTHROPOIETIN EXPRESSION BY DOXYCYCLINE IN RAT PRIMARY SKELETAL MUSCLE CELLS FOLLOWING TRANSDUCTION BY TETRACYCLINE GENE EXPRESSION SYSTEM". European Journal of General Medicine, vol. 3, no. 3, 2006, pp. 108-115. https://doi.org/10.29333/ejgm/82389

Electronic Journal of General Medicine

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