Dichomitus squalens, a promising white-rot basidiomycete for industrial enzyme production, necessitates efficient genetic manipulation systems to fully leverage its biotechnological potential. Established methodologies, including protoplast-mediated and Agrobacterium tumefaciens-mediated transformations, although functional in D. squalens, are intricate and time-consuming. This study introduces the electroporation transformation system for D. squalens, which is simpler and timesaving. By optimizing electroporation parameters, we obtained 77 transformants per μg of DNA. Furthermore, we confirmed the suitability of the Nourseothricin N-acetyl transferase gene as a selectable marker and the NanoLuc gene as a luciferase reporter in D. squalens using our refined electroporation protocol. This study expands the toolkit for genetic engineering in D. squalens, offering greater flexibility for future molecular investigations.