In this study, we investigated the design and construct of a chitosan‐based targeted gene delivery system and evaluated its function. To this end, chitosan‐folic acid/pDNA (CA‐FA/pDNA) nanoparticles were prepared in different formulations using the ion gelation method. All the synthesized nanoparticles were characterized using FTIR, TEM, SEM and DLS analysis. Moreover, the effects of molecular weight (MW) of chitosan (CA), DNA and CA concentration were inspected on encapsulation efficiency (EE). The results showed that the EE of pDNA was directly proportional with MW of CA and CA concentration but was in an inverse proportion with DNA concentration. In addition, HMwC and LMwC nanoparticles showed lower and higher pDNA release in all pH ranges, respectively. It is concluded that the N/P ratio increase can cause controlled pDNA release
