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

Koutroumpa FA, Monsempes C, Fran├žois 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

S. littoralis Orco (SlitOrco) gene structure and guide RNA positions.SlitOrco consists of 10 exons (grey boxes, E) and 9 introns. The number of amino acids encoded by each exon is indicated in the box. Two gRNAs, cr104 and cr105, were designed in exon 2 (E2) and one, cr106, in exon 4 (E4). Exon 2 and 4 sequences are detailed (underligned parts) showing positions of gRNAs (in blue ; PAM in italic and in box). Primers used for PCR amplification in the genotyping assay are in red italic.
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f1: S. littoralis Orco (SlitOrco) gene structure and guide RNA positions.SlitOrco consists of 10 exons (grey boxes, E) and 9 introns. The number of amino acids encoded by each exon is indicated in the box. Two gRNAs, cr104 and cr105, were designed in exon 2 (E2) and one, cr106, in exon 4 (E4). Exon 2 and 4 sequences are detailed (underligned parts) showing positions of gRNAs (in blue ; PAM in italic and in box). Primers used for PCR amplification in the genotyping assay are in red italic.

Mentions: 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. The gene structure consisted of 10 exons and 9 introns, comparable in size and positions with its B. mori orthologue (Fig. 1).


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

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

S. littoralis Orco (SlitOrco) gene structure and guide RNA positions.SlitOrco consists of 10 exons (grey boxes, E) and 9 introns. The number of amino acids encoded by each exon is indicated in the box. Two gRNAs, cr104 and cr105, were designed in exon 2 (E2) and one, cr106, in exon 4 (E4). Exon 2 and 4 sequences are detailed (underligned parts) showing positions of gRNAs (in blue ; PAM in italic and in box). Primers used for PCR amplification in the genotyping assay are in red italic.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: S. littoralis Orco (SlitOrco) gene structure and guide RNA positions.SlitOrco consists of 10 exons (grey boxes, E) and 9 introns. The number of amino acids encoded by each exon is indicated in the box. Two gRNAs, cr104 and cr105, were designed in exon 2 (E2) and one, cr106, in exon 4 (E4). Exon 2 and 4 sequences are detailed (underligned parts) showing positions of gRNAs (in blue ; PAM in italic and in box). Primers used for PCR amplification in the genotyping assay are in red italic.
Mentions: 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. The gene structure consisted of 10 exons and 9 introns, comparable in size and positions with its B. mori orthologue (Fig. 1).

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