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.