Conventional chemotherapy suffers lack of bioavailability, selectivity and multidrug resistance (MDR). Nano-sized drug delivery systems (DDS) is developing aimed to solve several limitations of conventional drug delivery systems. These systems have been offered for targeting tumor tissue due to their long circulation time and improved drug solubility, retention (EPR) effect, and enhanced permeability. So, the aim of this research was the development and design of a novel targeted nanocarrier for cancer chemotherapy. For this reason, chitosan (CS) was first modified with a chain transfer agent (CTA) to create CS-CTA macroinitiator. Then, the controlled grafting polymerization of itaconic acid (IA) and dimethylaminoethyl methacrylate quaternary ammonium alkyl halide (DMAEMAQ) monomers were occurred using reversible addition-fragmentation chain transfer (RAFT) polymerization to generate CS-g-(PIA-co-PDMAEMAQ) nanomicelles. To follow the cancer cells, fluorescein dye was entrapped into the core of nanomicelles and methotrexate (MTX) anticancer drug as a target ligand was incorporated into the cationic segment of nanomicelles. The chemical structures, biocompatibility, MTX loading capacity, in-vitro cytotoxicity effects and drug targeting ability of the developed nanomicelles were also investigated. Finally, it is expected that the nanomicelles can be used as a novel platform for targeted delivery.