Gene editing in Pyrrhocoris apterus established
The article in Frontiers in Physiology describes detail protocol for CRISPR-CAS9 gene editing in Pyrrhocoris apterus.
Description of the figure: Simplified workflow of the CRISPR/Cas9 mutant production in non-model insect Pyrrhocoris apterus with key optimization steps, and indicated duration of experiments. Note that the duration depends on generation time of particular insect species, yet, the necessary backcrossing to wild type is the most time-consuming part of the entire experiment. The optimal time for injections is species specific, however, in vivo sgRNA efficiency test as well as detection of G0 gonadal mosaicism can be easily adapted to any insect species.
The linden bug, Pyrrhocoris apterus, has served as an excellent model for research in insect endocrinology, chronobiology and developmental biology. However, reverse genetic tools were limited to RNA mediated interference. Therefore, we explored possibility to apply emerging gene editing technologies and optimized several steps needed for successful injection of embryos, efficient detection of mosaic founder animals and developed screening in live individuals of F1 generation. Our publication in Frontiers in Physiology describes the detail protocol for CRISPR-CAS9 gene editing Pyrrhocoris apterus, which can be, with certain modifications, applicable to other insect models.
Kotwica-Rolinska J., Chodakova L., Chvalova D., Kristofova L., Fenclova I., Provaznik J., Wu C., Bertolutti M., Dolezel D. (2019) CRISPR/Cas9 genome editing introduction and optimization in the non-model insect Pyrrhocoris apterus. Frontiers in Physiology 10: 891. DOI: 10.3389/fphys.2019.00891