Heritable genome editing with CRISPR/Cas9 induces anosmia in a crop pest moth.

Marie-Christine François,Emmanuelle Jacquin-Joly,Fotini A Koutroumpa,Christelle Monsempes,Corinne Royer,Jean-Paul Concordet,Anne De Cian

doi:10.1038/srep29620

Abstract

Lepidoptera suffer critical lack of genetic tools and heritable genome edition has been achieved only in a few model species. Here we demonstrate that the CRISPR/Cas9 system is highly efficient for genome editing in a non-model crop pest Lepidoptera, the noctuid moth Spodoptera littoralis. We knocked-out the olfactory receptor co-receptor Orco gene to investigate its function in Lepidoptera olfaction. We find that 89.6% of the injected individuals carried Orco mutations, 70% of which transmitted them to the next generation. CRISPR/Cas9-mediated Orco knockout caused defects in plant odor and sex pheromone olfactory detection in homozygous individuals. Our work genetically defines Orco as an essential OR partner for both host and mate detection in Lepidoptera, and demonstrates that CRISPR/Cas9 is a simple and highly efficient genome editing technique in noctuid pests opening new routes for gene function analysis and the development of novel pest control strategies.

Highlights

Lepidoptera suffer critical lack of genetic tools and heritable genome edition has been achieved only in a few model species
RNA interference (RNAi) approaches are usually inefficient in these species[1] and heritable mutagenesis has been established in only a limited number of model species
The genomic sequence of the S. littoralis Orco gene was identified via PCR, according to the B. mori Orco sequence (SilkDB: http://silkworm.genomics.org.cn/) assuming that intron positions would be similar

Summary

Introduction

Lepidoptera suffer critical lack of genetic tools and heritable genome edition has been achieved only in a few model species. ZFN-mediated mutagenesis has been recently reported for B. mori[4] and Danaus plexippus[5]; TALEN has been developed in B. mori[6,7,8]; and CRISPR/Cas[9] proved to be efficient in this species[9,10] as well as in the model butterflies Papilio xuthus[11] and Danaus plexippus[12]. The latter approach is in full expansion[13,14] and represents a groundbreaking milestone for non-model species. The present study aims at extending its use in non-model

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Results

Conclusion