Simple electroporation for efficient CRISPR/Cas9 genome editing in murine zygotes v4

Electroporation of zygotes represents a rapid alternative to the elaborate pronuclear injection procedure for CRISPR/Cas9-mediated genome editing in mice. However, current protocols for electroporation either require the investment in specialized electroporators or corrosive pre-treatment of zygotes which compromises embryo viability. Here, we describe an easily adaptable approach for highly efficient introduction of specific mutations in mice by electroporation of intact zygotes using a common electroporator with synthetic CRISPR/Cas9 components and minimal technical requirement. This protocol works efficiently with zygotes from a variety of genetic backgrounds and is compatible with other CRISPR nucleases like Cas12a.