Minos -mediated transgenesis in the pantry moth Plodia interpunctella

Transposon-mediated transgenesis has been widely used to study gene function in Lepidoptera, with piggyBac being the most commonly employed system. However, because the piggyBac transposase originates from a lepidopteran genome, concerns about potential endogenous activity and remobilization raise questions about its suitability for long-term genomic modifications across a wide range of species. As an alternative, we tested the effectiveness of the Minos transposase, a Tc1/mariner family element derived from a dipteran insect, for stable germline transformation in the pantry moth Plodia interpunctella . By injecting a Minos -based construct along with in vitro synthesized Minos transposase mRNA into syncytial embryos, we observed somatic activity of the 3xP3::EGFP eye/glia reporter gene in only 1 out of 14 (7.1%) injected G ₀ individuals that reached the pupal stage.

Nonetheless, pooled in-crossing of G ₀ individuals resulted in high rates of germline transmission of the transgene, with 23.6% of G ₁ pupae showing 3xP3 -driven eye fluorescence. Together with a previous study in Bombyx silkworms, these data suggest that Minos -mediated transgenesis yields practical rates of germline integration in Lepidoptera, providing an alternative tool for functional genomics in species where piggyBac may not be optimal.