The present investigation was carried out during summer 2017 and 2018 at El-Mattana Agricultural Research Station, Agricultural Research Center, Luxor, Egypt, to study combining ability and heterosis for yield and its components via line x tester analysis in sesame. Line × Tester mating design was applied using six lines and four testers and 24 F1 crosses evaluated in three replications of randomized block design. Results indicated that genotypes and parents exhibited highly significant differences for all studied traits, except parents for number of capsule per plant indicating that variability did exist among all entries. Parental genotypes line 51 and line 43 were the best combiner for seed yield per plant and its attributes. Besides, parental cultivar sohag-1 was a good combiner for plant height, seed yield per plant and seed oil content. Results showed that mean squares due to both general (GCA) and specific (SCA) combining ability were highly significant for all characters, indicating the importance of both additive and non-additive gene effects in the inheritance of the studied traits. Results also suggested that several crosses exhibited significant positive heterosis relative to both mid and better parents for all studied traits. Generally, three crosses, namely line 51 x Line F10-25, line 51 x Sohag-1 and line 43 x Sohag-1 had significant positive SCA and exceeded their mid and better parents for seed yield per plant, number of capsule per plant and oil content. These crosses could be exploited for breeding program to improving this crop.