Electroporation of mice zygotes with dual guide RNA/Cas9 complexes for simple and efficient cloning-free genome editing.
Sci Rep, 8(1):474.
In this report, we present an improved protocol for CRISPR/Cas9 genome editing in mice. The procedure consists in the electroporation of intact mouse zygotes withribonucleoprotein complexes prepared in vitro from recombinant Cas9 nuclease andsynthetic dual guide RNA. This simple cloning-free method proves to be extremelyefficient for the generation of indels and small deletions by non-homologous endjoining, and for the generation of specific point mutations by homology-directedrepair. The procedure, which avoids DNA construction, in vitro transcription andoocyte microinjection, greatly simplifies genome editing in mice.