CRISPR/Cas9-Based Gene Editing in Soybean

Methods Mol Biol. 2020:2107:349-364. doi: 10.1007/978-1-0716-0235-5_19.

Abstract

CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR associated Cas9)-based gene editing is a robust tool for functional genomics research and breeding programs in various crops. In soybean, a number of laboratories have obtained mutants by CRISPR/Cas9 system; however, there has been not yet a detailed method for the CRISPR/Cas9-based gene editing in soybean. Here, we describe the procedures for constructing the CRISPR/Cas9 plasmid suitable for soybean gene editing and the modified protocols for Agrobacterium-mediated soybean transformation and regeneration from cotyledonary node explants containing the Cas9/sgRNA (single guide RNA) transgenes.

Keywords: Agrobacterium-mediated soybean transformation; CRISPR/Cas9; Cotyledonary node; Gene editing; Soybean; Tissue culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium / genetics
  • CRISPR-Cas Systems
  • Cotyledon / genetics
  • Cotyledon / growth & development*
  • Gene Editing / methods*
  • Glycine max / genetics
  • Glycine max / growth & development*
  • Mutation
  • Plant Breeding
  • Tissue Culture Techniques
  • Transformation, Genetic