Highly efficient hyPBase-mediated transgenesis facilitates gain-of-function analysis in the cricket Gryllus...

Gain-of-function analysis is critical to directly assessing gene functions but remains challenging in non-traditional model insects. Recent development of the hyperactive piggyBac transposase (hyPBase) has improved the efficiency of germline transformation in ametabolous and holometabolous insects. Here, we report highly efficient somatic and germline transformations mediated by hyPBase in the cricket Gryllus bimaculatus. hyPBase-mediated misexpression of the melanin pathway gene arylalkylamine N-Acetyltransferase (aaNAT) turned the cricket body color from black to a light color, indicating a broad effect on external phenotype in the injected generation. Mosaic expression of the homeotic gene Antennapedia with hyPBase successfully circumvented the potential embryonic lethality, and induced antenna-to-leg homeotic transformation in a hatched nymph. Our data provide a practical methodology for gain-of-function analysis in the hemimetabolous model to understand key evolutionary innovations in insects.