Our technique: GEEP method 

 (Japanese Patent No. 6980218)

Genome Editing by Electroporation of Cas9 Protein (GEEP) method

Genome Editing by Electroporation of Cas9 Protein (GEEP) method is a method of introducing genome editing tools such as Cas9 protein and gRNA into fertilized eggs by the power of electricity (electroporation). The microinjection method, which has been used to introduce genome editing tools into fertilized eggs, requires specialized equipment such as a micromanipulator and advanced technique. On the other hand, by the GEEP method, all that is required is to line up the fertilized eggs on the electrode, and it is possible to introduce a genome editing tools into multiple (20 to 200) fertilized eggs at one time.

With GEEP method, we can produce genome-editing organisms highly efficiently. Also GEEP method can perform genome editing on a large number of fertilized eggs with minimal invasiveness under identical conditions in a short time.

The GEEP method is an innovative genome-editing method that enables genome editing with higher efficiency and quality than conventional methods.

Reference

  1. Hashimoto, M. and Takemoto, T*. Electroporation enables the efficient mRNA delivery into the mouse zygotes and facilitates CRISPR/Cas9-based genome editing. Sci. Rep. 5, 11315; doi: 10.1038/srep11315 (2015).
  2. Hashimoto M, Yamashita Y, Takemoto T*.Electroporation of Cas9 protein/sgRNA into early pronuclear zygotes generates non-mosaic mutants in the mouse. Dev. Biol. 418: 1-9 (2016).
  3. Tanihara F, Takemoto T*, Kitagawa E, Rao S, Do L, Onishi A, Yamashita Y, Kosugi C, Suzuki H, Sembon S, Suzuki S, Nakai M, Hashimoto M, Yasue A, Matsuhisa M, Noji N, Fujimura T, Fuchimoto Di, Otoi T*. Somatic cell reprogramming-free generation of genetically modified pigs. Science Advances. 2 (9) e1600803 (2016).