Mouse models with genetic modifications are widely used in biology and biomedical research. Although the application of CRISPR-Cas9 system greatly accelerated the process of generating genetically modified mice, the delivery method depending on manual injection of the components into the embryos remains a bottleneck, as it is laborious, low throughput, and technically demanding. To overcome this limitation, we invented and optimized the ZEN (Zygote electroporation of nucleases) technology to deliver CRISPR-Cas9 reagents via electroporation. Using ZEN, we were able to generate genetically modified mouse models with high efficiency and throughput. Here, we describe the protocol in great detail.
Keywords: CRISPR-Cas9; Electroporation; Mouse model generation; Mouse zygotes; ZEN.