Insects have the phenomenon of immune priming by which they can have enhanced protection against reinfection with the same pathogen, and this immune protection can be passed on to their offspring, which is defined as “trans-generational immune priming (TGIP).” But whether housefly possesses TGIP is still unclear. Therefore, we used the housefly as the insect model and Candida albicans as the pathogen to explore whether the housefly is capable of eliciting TGIP, and RNA sequencing (RNA-seq) was performed to explore the molecular mechanism of TGIP of the housefly. We found that the housefly possesses TGIP, and adults pre-exposed to heat-killed C. albicans could confer protection to itself and its offspring upon reinfection with a lethal dose of C. albicans. RNA-seq results showed that 30 and 154 genes were differentially expressed after adults were primed with heat-killed C. albicans (CA-A) and after offspring larvae were challenged with a lethal dose of C. albicans (CA-CA-G), respectively. Among the differentially expressed genes (DEGs), there were 23 immune genes, including 6 pattern recognition receptors (PRRs), 7 immune effectors, and 10 immunoregulatory molecules. More importantly, multiple DEGs were involved in the Toll signaling pathway and phagosome signaling pathway, suggesting that the Toll signaling pathway and phagocytosis might play important roles in the process of TGIP of housefly to C. albicans. Our results expanded on previous studies and provided parameters for exploring the mechanism of TGIP.