Genome Editing in Higher Plants

Abstract

Genome editing with engineered nucleases (“GEEN”) has emerged as an effective genetic engineering method that uses ‘molecular scissors’—artificially engineered nucleases—to digest DNA at targeted locations in the genome of various organisms including plant species. The DNA binding domains of zinc finger (ZF) proteins were first used as plant genome editing tools via the use of designed ZF nucleases (ZFNs), with TAL-effectors (TALE) and the RNA-DNA recognition system CRISPR/Cas9 now being used as powerful genome editing tools to create targeted gene modifications, not only in model plants but also in crop species. The key to genome editing is the introduction of targeted gene-specific double-stranded DNA breaks (DSBs) using the designed endonucleases, then allowing site-directed mutagenesis via nonhomologous end joining (NHEJ) repair and/or gene targeting via homologous recombination (HR), to occur efficiently at specific sites in the genome. This chapter provides an overview of recent advances in genome editing technologies, giving an insight into current plant molecular biology and breeding techniques.