(A) Location of the 10 bp deletion induced in the first exon of the SlitOr5 gene by the CRISPR/Cas9 system. The sequence complementary to the RNA guide is indicated in blue, and the protospacer adjacent motif (PAM) in red. The frameshift created in the SlitOr5 open-reading frame (ORF) induced a premature stop codon. (B) Electroantennogram (EAG) amplitude measured in S. littoralis male antennae isolated from wild-type animals (light grey, n = 14), heterozygous SlitOr5 mutants (dark grey, n = 18) and homozygous SlitOr5 mutants (purple, n = 8) after stimulation with pheromone compounds (1 µg in the stimulus cartridge) and plant volatiles (10 µg in the stimulus cartridge). Plotted values represent the normalized mean response ± SEM (response to the solvent was subtracted). ***p<0.001, significantly different from the response of the other genotypes; n.s.: not significantly different (one-way ANOVA, followed by a Tukey’s post hoc test). Raw results for the EAG experiment are available in Figure 3—source data 1. (C) Cumulative proportion of S. littoralis males initiating a movement toward the odor source in homozygous SlitOr5 mutants (purple, n = 14) stimulated with the major pheromone component (100 ng in the stimulus cartridge) and in wild-type animals stimulated with the pheromone (blue, n = 36) or with solvent alone (light grey, n = 44). ***p<0.001, significantly different from the other distributions; n.s.: not significantly different (log-rank test). Results obtained for other behavioral items are presented in Figure 3—figure supplement 1. (D) Proportion of wild-type S. littoralis males (light grey, n = 49) and homozygous SlitOr5 mutants (purple, n = 13) that mated with a wild-type female during a period of 6 hr in the scotophase. *p<0.05, significant difference between the two genotypes (Fisher’s exact test). (E) Number of eggs laid (left panel) and of offspring (right panel) obtained per female after the mating experiment. Plotted values represent the mean ± SEM. ***p<0.001, **p<0.005, significant difference between the two genotypes (Mann-Whitney U test). Raw results for all the behavioral experiments are available in Figure 3—source data 2.