The sonication-assisted whisker method enables CRISPR-Cas9 ribonucleoprotein delivery to induce genome editing in rice

Autor: Akiyoshi Nakamura, Tsubasa Yano, Nobutaka Mitsuda, Maiko Furubayashi, Seiichiro Ito, Shigeo S. Sugano, Teruhiko Terakawa
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: Scientific Reports, Vol 13, Iss 1, Pp 1-10 (2023)
Druh dokumentu: article
ISSN: 2045-2322
67940889
DOI: 10.1038/s41598-023-40433-w
Popis: Abstract CRISPR/Cas9-based genome editing represents an unprecedented potential for plant breeding. Unlike animal cells, plant cells contain a rigid cell wall, genome editing tool delivery into plant cells is thus challenging. In particular, the delivery of the Cas9-gRNA ribonucleoprotein (RNP) into plant cells is desired since the transgene insertion into the genome should be avoided for industrial applications in plants. In this study, we present a novel RNP delivery approach in rice. We applied the sonication-assisted whisker method, conventionally developed for DNA delivery in plants, for RNP delivery in rice. Combined with marker gene delivery, we successfully isolated Os LCYβ genome-edited lines generated by RNPs. The calli and regenerated shoot of the Os LCYβ mutant showed abnormal carotenoid accumulation. In addition, we also detected, although at a low frequency, genome editing events in rice calli cells by RNP delivery using the sonication-assisted whisker method without any additional. Therefore, the sonication-assisted whisker method could be an attractive way to create RNP-based genome-edited lines in plants.
Databáze: Directory of Open Access Journals
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