UNI-MB - logo
UMNIK - logo
 
E-viri
Recenzirano Odprti dostop
  • An optimized electroporatio...
    Tröder, Simon E; Ebert, Lena K; Butt, Linus; Assenmacher, Sonja; Schermer, Bernhard; Zevnik, Branko

    PloS one, 05/2018, Letnik: 13, Številka: 5
    Journal 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 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 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.