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

Koutroumpa FA, Monsempes C, François MC, de Cian A, Royer C, Concordet JP, Jacquin-Joly E - Sci Rep (2016)

Bottom Line: Lepidoptera suffer critical lack of genetic tools and heritable genome edition has been achieved only in a few model species.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.

View Article: PubMed Central - PubMed

Affiliation: INRA, UMR iEES-Paris, route de Saint-Cyr, 78026 Versailles Cedex, France.

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.

No MeSH data available.


Related in: MedlinePlus

Electrophysiological impact of Orco KO in S. littoralis antennae.Electroantennogram responses (EAG, in mV ±SEM, the response to solvent was subtracted) of S. littoralis antennae isolated from wild-type (WT, non-injected parents) and CRISPR/Cas9 G2 individuals toward plant odorants (10 μg), the main pheromone component (Z9,E11-14:Ac, 1 μg) and propionic acid (10 μg). (a) Female responses: WT (black), +/+ (grey, CRISPR/Cas9 G2 without any mutation), +/− (pink, CRISPR/Cas9 G2 with heterozygous mutation) and −/− (red, CRISPR/Cas9 G2 with homozygous mutation) (sequences 5, 15 and 16 as in Fig. 3, causing a truncated Orco protein); (b) Male responses as in (a); (c) Responses from males generated from parents carrying Orco sequence 1 (2 amino acid loss compared to the Orco wild-type sequence, no stop codon produced). Different letters above each odorant response indicate significant differences (t-test; p < 0.001); a is different from b and ab is not different from either a or b. PAA: 2-phenyl acetaldehyde; Ac: acetate ; OH: alcohol; n: number of individuals tested for each genotype.
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f4: Electrophysiological impact of Orco KO in S. littoralis antennae.Electroantennogram responses (EAG, in mV ±SEM, the response to solvent was subtracted) of S. littoralis antennae isolated from wild-type (WT, non-injected parents) and CRISPR/Cas9 G2 individuals toward plant odorants (10 μg), the main pheromone component (Z9,E11-14:Ac, 1 μg) and propionic acid (10 μg). (a) Female responses: WT (black), +/+ (grey, CRISPR/Cas9 G2 without any mutation), +/− (pink, CRISPR/Cas9 G2 with heterozygous mutation) and −/− (red, CRISPR/Cas9 G2 with homozygous mutation) (sequences 5, 15 and 16 as in Fig. 3, causing a truncated Orco protein); (b) Male responses as in (a); (c) Responses from males generated from parents carrying Orco sequence 1 (2 amino acid loss compared to the Orco wild-type sequence, no stop codon produced). Different letters above each odorant response indicate significant differences (t-test; p < 0.001); a is different from b and ab is not different from either a or b. PAA: 2-phenyl acetaldehyde; Ac: acetate ; OH: alcohol; n: number of individuals tested for each genotype.

Mentions: Non-mutated G2 (−/−) presented similar EAG responses as wild type adults when stimulated with plant-odorants and the main pheromone component32. Furthermore, we evidenced propionic acid detection in both wild type and −/− G2 males and females. The heterozygotes responded to all odorants with no statistically significant difference from the wild-types (p > 0,001, Fig. 4a,b), except for E-ocimen that induced very low EAG responses.


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

Koutroumpa FA, Monsempes C, François MC, de Cian A, Royer C, Concordet JP, Jacquin-Joly E - Sci Rep (2016)

Electrophysiological impact of Orco KO in S. littoralis antennae.Electroantennogram responses (EAG, in mV ±SEM, the response to solvent was subtracted) of S. littoralis antennae isolated from wild-type (WT, non-injected parents) and CRISPR/Cas9 G2 individuals toward plant odorants (10 μg), the main pheromone component (Z9,E11-14:Ac, 1 μg) and propionic acid (10 μg). (a) Female responses: WT (black), +/+ (grey, CRISPR/Cas9 G2 without any mutation), +/− (pink, CRISPR/Cas9 G2 with heterozygous mutation) and −/− (red, CRISPR/Cas9 G2 with homozygous mutation) (sequences 5, 15 and 16 as in Fig. 3, causing a truncated Orco protein); (b) Male responses as in (a); (c) Responses from males generated from parents carrying Orco sequence 1 (2 amino acid loss compared to the Orco wild-type sequence, no stop codon produced). Different letters above each odorant response indicate significant differences (t-test; p < 0.001); a is different from b and ab is not different from either a or b. PAA: 2-phenyl acetaldehyde; Ac: acetate ; OH: alcohol; n: number of individuals tested for each genotype.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940732&req=5

f4: Electrophysiological impact of Orco KO in S. littoralis antennae.Electroantennogram responses (EAG, in mV ±SEM, the response to solvent was subtracted) of S. littoralis antennae isolated from wild-type (WT, non-injected parents) and CRISPR/Cas9 G2 individuals toward plant odorants (10 μg), the main pheromone component (Z9,E11-14:Ac, 1 μg) and propionic acid (10 μg). (a) Female responses: WT (black), +/+ (grey, CRISPR/Cas9 G2 without any mutation), +/− (pink, CRISPR/Cas9 G2 with heterozygous mutation) and −/− (red, CRISPR/Cas9 G2 with homozygous mutation) (sequences 5, 15 and 16 as in Fig. 3, causing a truncated Orco protein); (b) Male responses as in (a); (c) Responses from males generated from parents carrying Orco sequence 1 (2 amino acid loss compared to the Orco wild-type sequence, no stop codon produced). Different letters above each odorant response indicate significant differences (t-test; p < 0.001); a is different from b and ab is not different from either a or b. PAA: 2-phenyl acetaldehyde; Ac: acetate ; OH: alcohol; n: number of individuals tested for each genotype.
Mentions: Non-mutated G2 (−/−) presented similar EAG responses as wild type adults when stimulated with plant-odorants and the main pheromone component32. Furthermore, we evidenced propionic acid detection in both wild type and −/− G2 males and females. The heterozygotes responded to all odorants with no statistically significant difference from the wild-types (p > 0,001, Fig. 4a,b), except for E-ocimen that induced very low EAG responses.

Bottom Line: Lepidoptera suffer critical lack of genetic tools and heritable genome edition has been achieved only in a few model species.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.

View Article: PubMed Central - PubMed

Affiliation: INRA, UMR iEES-Paris, route de Saint-Cyr, 78026 Versailles Cedex, France.

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.

No MeSH data available.


Related in: MedlinePlus