Tissue- and time-directed electroporation of CAS9 protein–gRNA complexes in vivo yields efficient multigene knockout for studying gene function in regeneration

This study introduces a method for efficient gene knockout in axolotls using electroporation of CAS9 protein–guide RNA (gRNA) complexes. The method allows for temporally and spatially controlled gene knockout, making it valuable for studying gene function in regeneration. The technique involves injecting CAS9-gRNA complexes into specific tissues and then using electroporation to deliver them into cells, resulting in efficient gene knockout.

• 1
  Electroporation of CAS9 protein–gRNA complexes is more efficient than plasmid-based CRISPR systems for gene knockout.
• 2
  The method can be used for multiplex gene knockout, allowing for the simultaneous knockout of multiple genes.
• 3
  The electroporation method can be applied to various tissues, including spinal cord, skin, and limb mesenchyme.