@article{ab4c986a55834e2abe3505ecb4059023,
title = "Protocol for Genome Editing to Produce Multiple Mutants in Wheat",
abstract = "Here, we describe a protocol for producing multiple recessive mutants via genome editing in hexaploid wheat (Triticum aestivum) cv. Fielder. Using Agrobacterium-delivered CRISPR/Cas9 and three sub-genome-specific primer sets, all possible combinations of single, double, and triple transgene-free mutants can be generated. The technique for acceleration of generation advancement with embryo culture reduces time for mutant production. The mutants produced by this protocol can be used for the analysis of gene function and crop improvement. For complete details on the use and execution of this protocol, please refer to Abe et al. (2019).",
author = "Fumitaka Abe and Yuji Ishida and Hiroshi Hisano and Masaki Endo and Toshihiko Komari and Seiichi Toki and Kazuhiro Sato",
note = "Funding Information: We thank Y. Kamiya and K. Kawaura (Kihara Institute for Biological Research, Yokohama City University) for helpful discussion, Y. Watanuki and K. Fujino (Institute of Crop Science, NARO) for technical assistance, and M. Shikata and H. Maeshima (NARO) for making the video. This work was supported by the Cabinet Office, Government of Japan, Cross-ministerial Strategic Innovation Promotion Program (SIP), ?Technologies for creating next-generation agriculture, forestry and fisheries? (funding agency: Bio-oriented Technology Research Advancement Institution, NARO). This work was also supported by JSPS KAKENHI grant number 19H00943. The seed sample of cv. Fielder was provided by the National BioResource Project?Wheat, Japan. Conceptualization, F.A. T.K. S.T. and K.S.; Investigation, F.A. Y.I. H.H. and M.E.; Resources, H.H. and M.E.; Writing ? Original Draft, F.A. Y.I. H.H. M.E. and K.S.; Writing ? Review & Editing, F.A. Y.I. H.H. M.E. and K.S.; Funding Acquisition, F.A. H.H. T.K. S.T. and K.S. The authors have a patent associated with this protocol (Method for gene transfer into triticum plant using agrobacterium bacterium, and method for production of transgenic plant of triticum plant. Patent no. WO2011013764A1). Funding Information: We thank Y. Kamiya and K. Kawaura (Kihara Institute for Biological Research, Yokohama City University) for helpful discussion, Y. Watanuki and K. Fujino (Institute of Crop Science, NARO) for technical assistance, and M. Shikata and H. Maeshima (NARO) for making the video. This work was supported by the Cabinet Office, Government of Japan , Cross-ministerial Strategic Innovation Promotion Program (SIP), “Technologies for creating next-generation agriculture, forestry and fisheries” (funding agency: Bio-oriented Technology Research Advancement Institution , NARO). This work was also supported by JSPS KAKENHI grant number 19H00943 . The seed sample of cv. Fielder was provided by the National BioResource Project–Wheat, Japan. Publisher Copyright: {\textcopyright} 2020 The Author(s)",
year = "2020",
month = sep,
day = "18",
doi = "10.1016/j.xpro.2020.100053",
language = "English",
volume = "1",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "2",
}