A chimeric antigen receptor-expressing T cell that targets and kills CD33 expressing cancers such as acute myeloid leukemia (AML). The CAR construct works by using a novel anti-CD33 scFv region to enable T cell targeting of CD33 expressing cancer cells and T-cell activation through the incorporation of co-stimulator and intracellular signaling regions. The anti-CD33 CAR-T cell-mediated killing of CD33-expressing CHO cells was demonstrated for up to 4 days. CD33 is a transmembrane receptor of the SIGLEC family that is expressed on myeloid cells. It is an effective target for treating AML as shown by in vivo AML mouse model studies where anti-CD33 CAR-T cells reduced leukemic burden and improved survival. Also, the FDA recently approved Mylotarg, an antibody-drug conjugate for the treatment of CD33(+) AML.