CRISPR/LbCas12a-Mediated Genome Editing in Soybean

Methods Mol Biol. 2023:2653:39-52. doi: 10.1007/978-1-0716-3131-7_3.

Abstract

Currently methods for generating soybean edited lines are time-consuming, inefficient, and limited to certain genotypes. Here we describe a fast and highly efficient genome editing method based on CRISPR-Cas12a nuclease system in soybean. The method uses Agrobacterium-mediated transformation to deliver editing constructs and uses aadA or ALS genes as selectable marker. It only takes about 45 days to obtain greenhouse-ready edited plants at higher than 30% transformation efficiency and 50% editing rate. The method is applicable to other selectable markers including EPSPS and has low transgene chimera rate. The method is also genotype-flexible and has been applied to genome editing of several elite soybean varieties.

Keywords: CRISPR-LbCas12a; Fast transformation; Genotype-independent transformation; Plant selectable marker gene; Soybean genome editing; Soybean transformation.

MeSH terms

  • CRISPR-Cas Systems* / genetics
  • Endonucleases / genetics
  • Gene Editing* / methods
  • Genome, Plant / genetics
  • Glycine max / genetics
  • Glycine max / metabolism
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism

Substances

  • Endonucleases