Supramolecular nanofibers (SNFs) composed of low‐molecular‐weight π‐conjugated molecules exhibit attractive optical and electrical properties and are expected to be the next optoelectronic materials. In this work, five crystalline SNFs have been constructed from three dehydrobenzoannulene (DBA) derivatives. The DBAs were designed to assemble in one dimension in a strategy based on anisotropic crystal growth. The crystallinity of the SNFs allowed the molecular arrangements in the SNFs to be determined. Therefore the mechanism of construction and correlations between the molecular arrangements and optical and electrical properties could be considered. The results clearly indicate that the properties of the SNFs are affected by the chemical structures and molecular arrangements. Moreover, one of the SNFs exhibits a high charge‐carrier mobility (Σμ=0.61 cm2 V−1 s−1) because of its crystallinity and appropriate molecular arrangement. This systematic experimental study based on a proposed strategy has provided information for improving the electrical properties of SNFs. This strategy will lead to highly functional SNFs.