Evaluation ofPeregrinus maidis transformer-2as a target for CRISPR-based control

Abstract

ABSTRACTThe corn planthopper,Peregrinus maidis, is an economically important pest of corn and sorghum. Here we report the initial steps towards developing a CRISPR-based control method, precision guided sterile insect technique (pgSIT), for this hemipteran pest. Specifically, we evaluated the potential oftransformer-2(tra-2) as a target for sterilizing insects. First, we identifiedtra-2transcripts within ourP. maidistranscriptome database and performed RNA interference (RNAi) to confirm functional conservation. RNAi-mediated knockdown ofPmtra-2in nymphs transformed females into pseudomales with deformed ovipositors resembling male claspers. While males showed no overt difference in appearance, they were indeed sterile. Importantly, the results were similar to those observed in another planthopper,Nilaparvata lugens. We also used CRISPR/Cas9 genome editing to assess the impact oftra-2knockout in injectees. CRISPR-mediated knockout ofPmtra-2had lethal effects on embryos, and hence not many injectees reached adulthood. However, mosaic knockout ofPmtra-2did impact female and male fertility, which supports the use oftra-2as a target for pgSIT in this hemipteran species.