Abstract: Gene therapy is an emerging strategy for the treatment of cardiovascular disorders. However, the efficient vehicle for gene delivery into myocardium is inadequately available to treat cardiovascular disorders. Despite the fact that many non-invasive protocols are used in gene delivery, the in vivo gene transfer into the heart is less specific and toxic. Therefore, alternative safe targeting methods are needed to deliver the specific gene into myocardium. In the present study, we developed an efficient method of gene transfer using nonhypertrophic receptor binding domain of Cardiac isoform of Alpha 2 Macroglobulin (CA2M) into myocardium. CA2M is a serum marker protein, which is cardiac specific and induced during hypertrophy in rats. In vivo gene transfer by direct injection of the truncated forms of CA2M into myocardium was performed in rats. The expression study using western blotting and quantification-using sandwich ELISA have identified the Nonhypertrophic Receptor-Binding Domain (NhRBD) of CA2M. Thus, the NhRBD of CA2M may be used as a novel vehicle to deliver the therapeutic gene into myocardium.