Gene Therapy is a promising technology for curing orthopedic health problems. However, the use of viral vectors for permanent integration could cause non-specific integration and negative side effects. Transient transfection is sufficient for some health issues like bone defects and especially healing skeletal defects like non-union fractures. After curing the defect, further overexpression of transgene growth factors, is neither necessary nor desired. Transient transfection is usually achieved by transfection with plasmid vectors, harboring sequences of bacterial origin. These are necessary for the amplification process. Minicircle -, a trimmed, circular DNA, has huge advantages for gene therapy. The lack of redundant bacterial DNA like antibiotic resistance genes lowers the risk of immune response as well as improving efficiency and duration of in vitro transfection. In this study a minicircle containing an expression cassette for bone morphogenic protein 2 (BMP2), which has strong osteoinductive activity, was designed. The functionality and osteoinductivity was tested in vitro in mouse myoblast progenitor cells (C2C12). Test results confirm the functionality of the BMP2-minicircle and promise 30.9-fold increase in BMP2 expression level compared to novel BMP2 expressing plasmid. |