The potato tuber moth, Phthorimaea operculella (Zeller), is a notorious insect pest of potato incurring substantial yield losses in the field as well as in storage. Chemical control is difficult to exercise due to the latent feeding of the caterpillars and their ability to develop resistance against insecticides. One of the essential components of efficient insect-resistant management is using two or more different insecticidal genes in transgenic crops to effectively avoid and delay the resistance development in insect pests. Two constructs, namely DS-1 (cry3A + SN-19 genes) and DS-2 (OCII + SN-19 genes) in pCAMBIA1301 binary vector, were developed and were transformed in potato cultivars (Agria and Lady Olympia) via Agrobacterium-mediated transformation. The molecular analysis confirmed gene integration and expression of the introduced genes in transgenic plants. The insecticidal effects of incorporated genes in transgenic plants were assessed against 1st, 2nd, 3rd, and 4th instar potato tuber moth (PTM) larvae. The transgenic plants endured significantly high mortalities (100%) of larval stages of PTM within 72 h. Our results show that these transgenic potato plants have the potential to control populations of PTM and are also useful tools in managing PTM that would ultimately reduce the dependency on conventional chemical pesticides with potentially less or minimal hazards. These lines can also serve as an excellent source of germplasm for potato breeding program.