Strategies for the Targeted Modification of Plant Genomes

The ability to precisely alter DNA sequences in living cells makes possible detailed functional analysis of genes and genetic pathways.  In plants, targeted genome modification has applications ranging from understanding plant gene function to developing crops with new traits of value.  We will describe efficient methods for targeted modification of plant genomes using sequence-specific nucleases.  With zinc finger nucleases (ZFNs), TAL effector nucleases (TALENs), and CRISPR/Cas9 reagents, we have achieved targeted gene knockouts, replacements and insertions in a variety of plant species.  Current work is focused on optimizing delivery of nucleases and donor DNA molecules to plant cells to more efficiently achieve targeted genetic alterations.