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Targeted mutagenesis using CRISPR-Cas9 in the chelicerate herbivore Tetranychus urticae

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  • SuperPests (Innovative tools for rational control of the most difficult-to-manage pests (super pests) and the diseases they transmit)
  • POLYADAPT (Molecular-genetic mechanisms of extreme adaptation in a polyphagous agricultural pest)
Abstract
The use of CRISPR-Cas9 has revolutionized functional genetic work in many organisms, including more and more insect species. However, successful gene editing or genetic transformation has not yet been reported for chelicerates, the second largest group of terrestrial animals. Within this group, some mite and tick species are economically very important for agriculture and human health, and the availability of a gene-editing tool would be a significant advancement for the field. Here, we report on the use of CRISPR-Cas9 in the spider mite Tetranychus urticae. The ovary of virgin adult females was injected with a mix of Cas9 and sgRNAs targeting the phytoene desaturase gene. Natural mutants of this laterally transferred gene have previously shown an easy-to-score albino phenotype. Albino sons of injected virgin females were mated with wild-type females, and two independent transformed lines where created and further characterized. Albinism inherited as a recessive monogenic trait. Sequencing of the complete target-gene of both lines revealed two different lesions at expected locations near the PAM site in the target-gene. Both lines did not genetically complement each other in dedicated crosses, nor when crossed to a reference albino strain with a known genetic defect in the same gene. In conclusion, two independent mutagenesis events were induced in the spider mite T. urticae using CRISPR-Cas9, hereby providing proof-of-concept that CRISPR-Cas9 can be used to create gene knockouts in mites.
Keywords
Insect Science, Biochemistry, Molecular Biology, Chelicerata, Genome editing, CRISPR, Cas9 ribonucleoprotein (RNP), Acari, GERMLINE TRANSFORMATION, CAENORHABDITIS-ELEGANS, SALIVARY PROTEINS, RESISTANCE, GENE, RESPONSES, MITES, MUTATION, GENOME, MODE

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Citation

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MLA
Dermauw, Wannes, et al. “Targeted Mutagenesis Using CRISPR-Cas9 in the Chelicerate Herbivore Tetranychus Urticae.” INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, vol. 120, 2020, doi:10.1016/j.ibmb.2020.103347.
APA
Dermauw, W., Jonckheere, W., Riga, M., Livadaras, I., Vontas, J., & Van Leeuwen, T. (2020). Targeted mutagenesis using CRISPR-Cas9 in the chelicerate herbivore Tetranychus urticae. INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, 120. https://doi.org/10.1016/j.ibmb.2020.103347
Chicago author-date
Dermauw, Wannes, Wim Jonckheere, Maria Riga, Ioannis Livadaras, John Vontas, and Thomas Van Leeuwen. 2020. “Targeted Mutagenesis Using CRISPR-Cas9 in the Chelicerate Herbivore Tetranychus Urticae.” INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY 120. https://doi.org/10.1016/j.ibmb.2020.103347.
Chicago author-date (all authors)
Dermauw, Wannes, Wim Jonckheere, Maria Riga, Ioannis Livadaras, John Vontas, and Thomas Van Leeuwen. 2020. “Targeted Mutagenesis Using CRISPR-Cas9 in the Chelicerate Herbivore Tetranychus Urticae.” INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY 120. doi:10.1016/j.ibmb.2020.103347.
Vancouver
1.
Dermauw W, Jonckheere W, Riga M, Livadaras I, Vontas J, Van Leeuwen T. Targeted mutagenesis using CRISPR-Cas9 in the chelicerate herbivore Tetranychus urticae. INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY. 2020;120.
IEEE
[1]
W. Dermauw, W. Jonckheere, M. Riga, I. Livadaras, J. Vontas, and T. Van Leeuwen, “Targeted mutagenesis using CRISPR-Cas9 in the chelicerate herbivore Tetranychus urticae,” INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, vol. 120, 2020.
@article{8655069,
  abstract     = {The use of CRISPR-Cas9 has revolutionized functional genetic work in many organisms, including more and more insect species. However, successful gene editing or genetic transformation has not yet been reported for chelicerates, the second largest group of terrestrial animals. Within this group, some mite and tick species are economically very important for agriculture and human health, and the availability of a gene-editing tool would be a significant advancement for the field. Here, we report on the use of CRISPR-Cas9 in the spider mite Tetranychus urticae. The ovary of virgin adult females was injected with a mix of Cas9 and sgRNAs targeting the phytoene desaturase gene. Natural mutants of this laterally transferred gene have previously shown an easy-to-score albino phenotype. Albino sons of injected virgin females were mated with wild-type females, and two independent transformed lines where created and further characterized. Albinism inherited as a recessive monogenic trait. Sequencing of the complete target-gene of both lines revealed two different lesions at expected locations near the PAM site in the target-gene. Both lines did not genetically complement each other in dedicated crosses, nor when crossed to a reference albino strain with a known genetic defect in the same gene. In conclusion, two independent mutagenesis events were induced in the spider mite T. urticae using CRISPR-Cas9, hereby providing proof-of-concept that CRISPR-Cas9 can be used to create gene knockouts in mites.},
  articleno    = {103347},
  author       = {Dermauw, Wannes and Jonckheere, Wim and Riga, Maria and Livadaras, Ioannis and Vontas, John and Van Leeuwen, Thomas},
  issn         = {0965-1748},
  journal      = {INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY},
  keywords     = {Insect Science,Biochemistry,Molecular Biology,Chelicerata,Genome editing,CRISPR,Cas9 ribonucleoprotein (RNP),Acari,GERMLINE TRANSFORMATION,CAENORHABDITIS-ELEGANS,SALIVARY PROTEINS,RESISTANCE,GENE,RESPONSES,MITES,MUTATION,GENOME,MODE},
  language     = {eng},
  pages        = {9},
  title        = {Targeted mutagenesis using CRISPR-Cas9 in the chelicerate herbivore Tetranychus urticae},
  url          = {http://dx.doi.org/10.1016/j.ibmb.2020.103347},
  volume       = {120},
  year         = {2020},
}

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