National Agriculture and Food Research Organization (NARO), in collaboration with Chiba University and Tokyo Institute of Technology, has developed a new technology that introduces enzymes necessary for genome editing into plant cells by short-time exposure to atmospheric pressure plasma. In the conventional genome editing technology, introduction of exogenous DNA is required, but this new technology does not require removal of exogenous DNA. Therefore, it is considered to be more convenient and applicable to various plants, and it is expected to become a new tool for breed improvement.
Overview
A research group led by Ichiro Mitsuhara, Group Leader, NARO along with Yuki Yanagawa, a specially appointed researcher at Chiba University, and Akitoshi Okino, Associate Professor, Tokyo Institute of Technology, has developed a technology to edit plant genomes by introducing a genome-editing enzyme (complex of Cas9 protein and sgRNA) into plant cells by irradiating them with atmospheric pressure plasma controlled at a low temperature of about 25°C.
In the past, plant genome editing used gene recombination technology to temporarily introduce enzyme genes in the form of DNA. After genome editing with this technology, the introduced genome-editing enzyme gene becomes unnecessary, so it was necessary to remove the foreign DNA by methods such as self-pollination or crossing to obtain the next generation of plants.
The technology developed this time utilizes the technology developed in 2017 by NARO and the Tokyo Institute of Technology to introduce biopolymers into plant bodies by irradiating them with atmospheric pressure plasma and genome-editing enzymes (protein RNA complexes) are introduced into plant cells. In this technology, genome-editing enzymes are not introduced in the form of genes (DNA), but directly into plant cells in the form of proteins or RNA. Therefore, the removal of foreign genes, which was required in conventional genome editing technology, is no longer necessary. In addition, compared to technologies such as bombardment method and electroporation method, it has the advantage of being able to introduce a wide range of enzymes into the cell population of plasma-irradiated plant tissue while maintaining the shape of the plant body. Hence it will be possible to modify the plant genomes more efficiently.
It is expected that this technology will be used as a new tool for the development of plant varieties for which genome editing has been difficult until now, such as trees with long life cycles and vegetatively propagated crops.
Publication
Yuki Yanagawa, Yuma Suenaga, Yusuke Iijima, Masaki Endo, Naoko Sanada, Etsuko Katoh, Seiichi Toki, Akitoshi Okino, Ichiro Mitsuhara.(2023) Genome editing by introduction of Cas9/sgRNA into plant cells using temperature-controlled atmospheric pressure plasma. PLoS ONE 18: e0281767. https://doi.org/10.1371/journal.pone.0281767
