The advent of nanobiotechnology in the science has changed the methods of preventation and treatment in medicine. Gene therapy is one of the new advanced methods of prevention and treatment that is rapidly developing by this multidisciplinary science. Different mechanisms have been defined for gene therapy till now that everyone has its own benefits. Undoubtedly, the presence of an appropriate vector for safe and maximum transgene to the site of vaccination or treatment and its successful release while creating the lowest toxicity is the main factor for the success of any of these mechanisms. In this study, the construction and properties of biomolecular coated magnetic iron oxide nanoparticles with ascorbic acid modified chitosan was investigated, which included the features mentioned below. Nanoparticles were studied by Fourier Infrared Transformation(FT-IR) spectrometry, Alkaline titration, X-ray diffraction(XRD), Vibration Sample Magnetometer(VSM), UV Spectrophotometry, Electrophoresis, Zeta Potential Measurement, and Dynamic Light Scattering(DLS). They were also photographed using a Field Emission Scanning Electron Microscope(FESEM). FT-IR spectroscopy and titration confirmed the successful transcription of ascorbic acid with chitosan. The X-ray diffraction analysis showed that synthesises of both simple and coated magnetite types were done correctly. The results of VSM test showed superparamagnetism of these nanoparticles and the saturation magnetization of the carrier nanoparticles was obtained 46 emu/g. The loading, stability, and release of vectors were monitored by ultraviolet photometric spectroscopy and electrophoresis. loading of all plasmid DNA in carrier nanoparticles and the complete stability of these nanoparticles were confirmed. The release of plasmid DNA from nanoparticles was also successful. The measured values ??for zeta potential and the average hydrodynamic diameter of carrier nanoparticles were in a range that based on previous studies, the best cellular uptake happens for carrier nanoparticles. By analyzing and summarizing the results and comparing the results of other researchers in this field, these nanoparticles have successfully completed in vitro studies. It seems that these vectors have the ability to use in other gene therapy techniques and even in other nanobiotechnology fields.