| Title: | Targeted mutagenesis using CRISPR-Cas9 in the chelicerate herbivore Tetranychus urticae |
| Author(s): | Dermauw W; Jonckheere W; Riga M; Livadaras I; Vontas J; Van Leeuwen T; |
| Address: | "Laboratory of Agrozoology, Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000, Ghent, Belgium. Electronic address: wannes.dermauw@ugent.be. Laboratory of Agrozoology, Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000, Ghent, Belgium. Molecular Entomology Lab, Institute of Molecular Biology and Biotechnology (IMBB), Foundation for Research and Technology (FORTH), Nikolaou Plastira Street 100, 70013, Heraklion, Crete, Greece. Molecular Entomology Lab, Institute of Molecular Biology and Biotechnology (IMBB), Foundation for Research and Technology (FORTH), Nikolaou Plastira Street 100, 70013, Heraklion, Crete, Greece; Pesticide Science Laboratory, Department of Crop Science, Agricultural University of Athens, Iera Odos 75, 11855, Athens, Greece. Laboratory of Agrozoology, Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, 9000, Ghent, Belgium. Electronic address: thomas.vanleeuwen@ugent.be" |
| DOI: | 10.1016/j.ibmb.2020.103347 |
| ISSN/ISBN: | 1879-0240 (Electronic) 0965-1748 (Linking) |
| 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: | Animals;Animals Arthropod Proteins/*genetics/metabolism *CRISPR-Cas Systems *Gene Editing *Mutagenesis Oxidoreductases/*genetics/metabolism Tetranychidae/*genetics Acari Crispr Cas9 ribonucleoprotein (RNP) Chelicerata Genome editing; |
| Notes: | "MedlineDermauw, Wannes Jonckheere, Wim Riga, Maria Livadaras, Ioannis Vontas, John Van Leeuwen, Thomas eng Research Support, Non-U.S. Gov't England 2020/03/03 Insect Biochem Mol Biol. 2020 May; 120:103347. doi: 10.1016/j.ibmb.2020.103347. Epub 2020 Feb 27" |
