May 27, 2016
Streamlining and optimizing plant genome editing for environmental robustness
Tokushima University and RIKEN CSRS researchers have developed a CRISPR-Cas9 technology that streamlines and optimizes plant genomes that result in strong environmental response capabilities.
The group determined that truncated gRNA (tru-gRNA)–Cas9 was effective in plant cells and reduced off-target effects in mutant plants. They also found that the expression region of the promoter used to control Cas9 expression and the developmental stages greatly affected results.
Tru-gRNA Cas9 cassettes driven by promotors specific to high expression in the germ line were used to induce a targeted mutation in the OST2 gene in Arabidopsis and allow high-efficiency production of second-generation mutants that had lost their OST2 gene due to Mendelian inheritance. The OST2 mutants had enhanced stomatal closure, showing that it is possible to engineer changes that affect environmental stress responses using CRISPR-Cas9.
The use of this genome editing technology in Arabidopsis demonstrates that plant genome editing is possible for various research purposes and should facilitate applications in a wide variety of areas such as agricultural production and environmental control.
Scientific Reports doi:10.1038/srep26685
Y. Osakabe, T. Watanabe, S.S. Sugano, R. Ueta, R. Ishihara, K. Shinozaki, K. Osakabe,
"Optimization of CRISPR/Cas9 genome editing to modify abiotic stress responses in plants".
Gene Discovery Research Group