STAR Protocols (Sep 2020)

Protocol for Genome Editing to Produce Multiple Mutants in Wheat

  • Fumitaka Abe,
  • Yuji Ishida,
  • Hiroshi Hisano,
  • Masaki Endo,
  • Toshihiko Komari,
  • Seiichi Toki,
  • Kazuhiro Sato

Journal volume & issue
Vol. 1, no. 2
p. 100053

Abstract

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Summary: 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).