Egg Microinjection and Efficient Mating for Genome Editing in the Firebrat Thermobia domestica

• Published in: Journal of visualized experiments no. 164
• Main Authors: Ohde, Takahiro, Minemura, Toshinori, Hirose, Eiichi, Daimon, Takaaki
• Format: Journal Article
• Language: English
• Published: United States 20.10.2020
• Subjects: Animals; Gene Editing - methods; Insecta - genetics; Microinjections; Ovum - physiology; Reproduction
• ISSN: 1940-087X; 1940-087X
• DOI: 10.3791/61885

The firebrat Thermobia domestica is an ametabolous, wingless species that is suitable for studying the developmental mechanisms of insects that led to their successful evolutionary radiation on the earth. The application of genetic tools such as genome editing is the key to understanding genetic changes that are responsible for evolutionary transitions in an Evo-Devo approach. In this article, we describe our current protocol for generating and maintaining mutant strains of T. domestica. We report a dry injection method, as an alternative to the reported wet injection method, that allows us to obtain stably high survival rates in injected embryos. We also report an optimized environment setting to mate adults and obtain subsequent generations with high efficiency. Our method underlines the importance of taking each species' unique biology into account for the successful application of genome editing methods to non-traditional model organisms. We predict that these genome editing protocols will help in implementing T. domestica as a laboratory model and to further accelerate the development and application of useful genetic tools in this species.