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.

Blog Article

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 the introduction Toilet Brushes of specific mutations in C57BL/6 mice by electroporation of intact zygotes using a Accessory (Set of 5) common electroporator with synthetic CRISPR/Cas9 components and minimal technical requirement.Direct comparison to conventional 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.