Loss-of-function in testis-specific serine/threonine protein kinase (TSSKs) triggers male infertility in an invasive moth

Abstract Genetic control system at molecular level presents a promising and eco-friendly strategy for the management of pest and insect-transmitted diseases. Although considerable advancements have been achieved in gene drive applications targeting mosquitoes, endeavors to combat agricultural pests have been somewhat restricted. Here, we identified that the testis-specific serine/threonine kinases (TSSKs) family is uniquely expressed in the testes of Cydia pomonella, a prominent global invasive species. We further generated male moths with disrupted the expression of TSSKs and those with TSSKs completely knocked out using RNA interference and CRISPR/Cas 9 genetic editing techniques, resulting in significant disruptions in spermiogenesis, decreased sperm motility, and hindered development of fertilized oocytes. Further explorations into the underlying post-transcriptional regulatory mechanisms have revealed the involvement of lnc117962 as a competing endogenous RNA (ceRNA) for miR-3960, thereby regulating TSSKs. Notably, orchard trials have demonstrated that the release of male strains can effectively suppress population growth. Our findings indicate that targeting TSSKs could serve as a feasible avenue for managing C. pomonella populations, offering significant insights and potential strategies for controlling invasive pests through genetic sterile insect technique (gSIT) technology.