Non-transgenic Plant Genome Editing Using Purified Sequence-Specific Nucleases

Sequence-specific nucleases, including zinc-finger nucleases, meganucleases, TAL effector nucleases (TALENs), and CRISPR/Cas systems, have been used to introduce targeted mutations in a wide range of plant species (Voytas, 2013; Baltes and Voytas, 2015). However, delivery of these nucleases using traditional transformation methods (e.g., particle bombardment, Agrobacterium or protoplast transformation) may result in undesired genetic alterations due to random insertion of nuclease-encoding DNA into the host genome.