An optimized electroporation approach for efficient CRISPR/Cas9 genome editing in murine zygotes.

An optimized electroporation approach for efficient CRISPR/Cas9 genome editing in murine zygotes.

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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 bunny spa headband or corrosive pre-treatment of zygotes which compromises embryo viability.Here, we describe an easily adaptable approach for the introduction of specific mutations in C57BL/6 mice by electroporation of intact zygotes using a common electroporator with synthetic CRISPR/Cas9 components and minimal technical requirement.Direct comparison to conventional cnd be demure pronuclear injection demonstrates significantly reduced physical damage and thus improved embryo development with successful genome editing in up to 100% of living offspring.Hence, our novel approach for Easy Electroporation of Zygotes (EEZy) allows highly efficient generation of CRISPR/Cas9 transgenic mice while reducing the numbers of animals required.