A DNA-free and genotype-independent CRISPR/Cas9 system in soybean.

Autor:	Kuwabara C; Graduate School of Agriculture, Hokkaido University, Kita 9, Nishi 9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan., Miki R; Agri-Bio Research Center, Kaneka Corporation, 700 Higashibara, Iwata, Shizuoka 438-0802, Japan., Maruyama N; Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan., Yasui M; Research Faculty of Agriculture, Hokkaido University, Kita 9, Nishi 9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan., Hamada H; Agri-Bio Research Center, Kaneka Corporation, 700 Higashibara, Iwata, Shizuoka 438-0802, Japan., Nagira Y; Agri-Bio Research Center, Kaneka Corporation, 700 Higashibara, Iwata, Shizuoka 438-0802, Japan., Hirayama Y; Genome-Edited Crop Development Group, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Kannondai 3-1-3, Tsukuba, Ibaraki 305-8604, Japan., Ackley W; Genome-Edited Crop Development Group, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Kannondai 3-1-3, Tsukuba, Ibaraki 305-8604, Japan., Li F; Division of Crop Design Research, Institute of Crop Science, National Agricultural and Food Research Organization (NARO), Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan., Imai R; Genome-Edited Crop Development Group, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Kannondai 3-1-3, Tsukuba, Ibaraki 305-8604, Japan.; Faculty of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan., Taoka N; Agri-Bio Research Center, Kaneka Corporation, 700 Higashibara, Iwata, Shizuoka 438-0802, Japan., Yamada T; Graduate School of Agriculture, Hokkaido University, Kita 9, Nishi 9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan.
Jazyk:	angličtina
Zdroj:	Plant physiology [Plant Physiol] 2024 Dec 02; Vol. 196 (4), pp. 2320-2329.
DOI:	10.1093/plphys/kiae491
Abstrakt:	Here, we report a smart genome editing system for soybean (Glycine max) using the in planta bombardment-ribonucleoprotein (iPB-RNP) method without introducing foreign DNA or requiring traditional tissue culture processes such as embryogenesis and organogenesis. Shoot apical meristem (SAM) of embryonic axes was used as the target tissue for genome editing because the SAM in soybean mature seeds has stem cells and specific cell layers that develop germ cells during the reproductive growth stage. In the iPB-RNP method, the RNP complex of the CRISPR/Cas9 system was directly delivered into SAM stem cells via particle bombardment, and genome-edited plants were generated from these SAMs. Soybean allergenic gene Gly m Bd 30K was targeted in this study. Many E0 (the first generation of genome-edited) plants in this experiment harbored mutant alleles at the targeted locus. Editing frequency of inducing mutations transmissible to the E1 generation was approximately 0.4% to 4.6% of all E0 plants utilized in various soybean varieties. Furthermore, simultaneous mutagenesis by iPB-RNP method was also successfully performed at other loci. Our results offer a practical approach for both plant regeneration and DNA-free genome editing achieved by delivering RNP into the SAM of dicotyledonous plants. Competing Interests: Conflict of interest statement. R.M., H.H., Y.N., and N.T. were employed by Kaneka Corporation. T.Y. receives research support from Kaneka Corporation. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)
Databáze:	MEDLINE
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