Gene therapy is a good alternative to drugs in chemotherapy due to drug resistance and toxicity and known to lower the probability of mutation of cells through gene carrier. So we aimed to manufacture a gene carrier with minimal toxicity and high efficiency. Biodegradable and biocompatible polymer LTU (L-tyrosine polyurethane) was synthesized using desaminotyrosyl tyrosine hexyl ester (DTH), polyethylene glycol (PEG) and the LTU (L-tyrosine polyurethane) nanoparticles were manufactured through double emulsion method. In order to study cell adsorption and transfection of nanoparticles, two types of nanoparticles were prepared. One nanoparticles was fluoresced using fluorescently labeled bovine serum albumin (FITC-BSA) to investigate cell adsorption, and the other nanoparticles encapsulated the DNA-linear polyethylenimine (LPEI) complex to investigate the transfection efficiency in LX2 (human hepatic stellate cell), HepG2 (human liver cancer cell). The morphology of these nanoparticles were confirmed by microscopy and TEM, which is a spherical form with an average size of about 260nm and 80nm. The LTU nanoparticles showed the typical features of double emulsion. After 14 days, 15 ng of DNA per mg of nanoparticles have been released from the LTU nanoparticles. Furthermore, a successful uptake of LTU nanoparticles by the hepatic cells has been confirmed with stability state. These characteristics are ideal for gene therapy designed to transport and release a drug into the cytoplasm.