Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation

Genome Biol. 2015 Jul 21;16(1):144. doi: 10.1186/s13059-015-0715-0.

Abstract

Arabidopsis mutants produced by constitutive overexpression of the CRISPR/Cas9 genome editing system are usually mosaics in the T1 generation. In this study, we used egg cell-specific promoters to drive the expression of Cas9 and obtained non-mosaic T1 mutants for multiple target genes with high efficiency. Comparisons of 12 combinations of eight promoters and two terminators found that the efficiency of the egg cell-specific promoter-controlled CRISPR/Cas9 system depended on the presence of a suitable terminator, and the composite promoter generated by fusing two egg cell-specific promoters resulted in much higher efficiency of mutation in the T1 generation compared with the single promoters.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / embryology
  • Arabidopsis / genetics*
  • CRISPR-Associated Proteins / genetics
  • CRISPR-Associated Proteins / metabolism
  • CRISPR-Cas Systems*
  • Genes, Plant*
  • Germ Cells, Plant / metabolism
  • Homozygote*
  • Mutation*
  • Phenotype
  • Promoter Regions, Genetic*
  • Terminator Regions, Genetic

Substances

  • CRISPR-Associated Proteins