Find in Library
Search millions of books, articles, and more
Indexed Open Access Databases
An optimized electroporation approach for efficient CRISPR/Cas9 genome editing in murine zygotes.
oleh: Simon E Tröder, Lena K Ebert, Linus Butt, Sonja Assenmacher, Bernhard Schermer, Branko Zevnik
Format: | Article |
---|---|
Diterbitkan: | Public Library of Science (PLoS) 2018-01-01 |
Deskripsi
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.