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TALENs and CRISPR/Cas9: Genome editing tools for wheat.
J. GIL-HUMANES (1), N. J. BALTES (1), D. F. VOYTAS (1). (1) UNIVERSITY OF MINNESOTA, St. Paul, MN, U.S.A.
TALENs have emerged as one of the most versatile methods for genome engineering in different organisms. These customizable molecular DNA scissors are capable of inducing double-strand breaks at given genomic locations, consequently producing mutations such as insertions, replacements or chromosome rearrangements after repair of the break. Due to the simplicity of the DNA recognition code and their modular assembly, TALENs can be designed to target virtually any DNA sequence. Thus, they provide an efficient approach to targeted genome editing of plants and other organisms. In the last years TALEN-mediated genome modification has been reported in a number of organisms including plants. In addition, the type II bacterial CRISPR/Cas system has also arisen as a facile and efficient platform for genome editing. Bacteria and archaea have evolved an RNA-based adaptive immune system that uses CRISPR (clustered, regularly interspaced, short palindromic repeat) and Cas (CRISPR-associated) proteins to detect and destroy invading viruses and plasmids. The system has been recently adapted to produce directed mutations in plant species. These cutting edge technologies will revolutionize genome engineering in plants, and offer new alternatives for genome editing of polyploid species such as wheat. Directed mutagenesis (gene knockouts, gene deletions, directed amino acid changes) and gene targeting into the desired locus are some of the possibilities that TALENs and CRISPR/Cas9 technologies offer for genome engineering of wheat. The enhancement of the mutation efficiency and the reduction of the off-target mutations of unrelated loci, as well as the development of delivery systems that do not involve transgene integration are some of the challenges that these technologies will be facing in the upcoming years.
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