Toward a CRISPR-Cas9-based Gene Drive in the Diamondback Moth Plutella xylostella

Promising to provide powerful genetic control tools, gene drives have been constructed in multiple dipteran insects, yeast, and mice for the purposes of population elimination or modification. However, it remains unclear whether these techniques can be applied to lepidopterans. Here, we used endogenous regulatory elements to drive Cas9 and single guide RNA (sgRNA) expression in the diamondback moth (DBM), Plutella xylostella, and test the first split gene drive system in a lepidopteran. The DBM is an economically important global agriculture pest of cruciferous crops and has developed severe resistance to various insecticides, making it a prime candidate for such novel control strategy development. A very high level of somatic editing was observed in Cas9/sgRNA transheterozygotes, although no significant homing was revealed in the subsequent generation. Although heritable Cas9-medated germline cleavage as well as maternal and paternal Cas9 deposition were observed, rates were far lower than for somatic cleavage events, indicating robust somatic but limited germline activity of Cas9/sgRNA under the control of selected regulatory elements. Our results provide valuable experience, paving the way for future construction of gene drives or other Cas9-based genetic control strategies in DBM and other lepidopterans.

Automated summary

This study successfully used endogenous regulatory elements to drive the expression of Cas9 and sgRNA in the diamondback moth (DBM), testing the first split gene drive system in a lepidopteran. The results showed high levels of somatic editing, but limited effects in germline cells. This research provides valuable insights for future development of gene drive systems or other Cas9-based genetic control strategies in DBM and other lepidopterans.