Eggplant shoot and fruit borer (ESFB), Leucinodes orbonalis Guenee, is a major insect pest of eggplant, Solanum melongena L. In this study, cry1Aa3 gene was introduced into eggplant cv. Kashi Taru through Agrobacterium-tumefaciens mediated transformation. Hypocotyl explants from 7 day-old seedlings were pre-cultured for two days and inoculated with Agrobacterium strain EHA105 harboring a synthetic cry1Aa3 gene with full codon-modification in a binary vector. PCR and Southern blot analyses confirmed the presence of single copy insertions of the cry1Aa3 gene in plants of four independent transformation events. In addition, single copy gene insertions were confirmed by segregation analysis of T1 seedlings. Expression of the cry1Aa3 transcripts was confirmed by qRT-PCR; and ELISA revealed presence of Cry1Aa3 protein in fresh leaves (30.9–44.3 ng g−1) and fruits (20.5–35.7 ng g−1). Expression of the Cry1Aa3 protein resulted in high ESFB larval mortality in the shoot as well as fruit tissues of the transformed eggplant lines. Such transgenic eggplant lines possessing resistance to the ESFB could reduce dependency on pesticide use for ESFB control, leading to a safer environment and eggplant fruits.