The binding and fluorescence properties of complementary RNA sequences attached to different numbers of pyrenes via one carbon linker at the 2′-O-positions have been investigated. Upon hybridization of the pyrene-modified RNA sequences, the modified RNA duplexes with normal thermal stability are formed, and the pyrene arrays are assembled in an inter-strand manner. Because hypochromic effects in the pyrene absorption band and the exciton coupled circular dichroism signals were observed for the pyrene assemblies, the formation of the pyrene array occurs via a π-stacking interaction between the pyrene rings. The pyrene assemblies exhibit strong excimer fluorescence that is characterized by a broad and structureless excitation spectrum. Hence, the excimer is a static excimer due to the direct excitation of the associated pyrenes in the ground state. Based on several spectroscopies, it is revealed that the spatial configuration of the pyrenes in the association is more regulated by the increase in the attached pyrene.