Genome editing in maize directed by CRISPR-Cas9 ribonucleoprotein complexes

Nat Commun. 2016 Nov 16:7:13274. doi: 10.1038/ncomms13274.

Abstract

Targeted DNA double-strand breaks have been shown to significantly increase the frequency and precision of genome editing. In the past two decades, several double-strand break technologies have been developed. CRISPR-Cas9 has quickly become the technology of choice for genome editing due to its simplicity, efficiency and versatility. Currently, genome editing in plants primarily relies on delivering double-strand break reagents in the form of DNA vectors. Here we report biolistic delivery of pre-assembled Cas9-gRNA ribonucleoproteins into maize embryo cells and regeneration of plants with both mutated and edited alleles. Using this method of delivery, we also demonstrate DNA- and selectable marker-free gene mutagenesis in maize and recovery of plants with mutated alleles at high frequencies. These results open new opportunities to accelerate breeding practices in a wide variety of crop species.

MeSH terms

  • Alleles
  • Base Sequence
  • Biolistics
  • CRISPR-Cas Systems / genetics*
  • DNA, Plant / metabolism
  • Gene Editing*
  • Gene Transfer Techniques
  • Genes, Plant
  • Multiprotein Complexes / metabolism*
  • Mutagenesis / genetics
  • Mutation / genetics
  • Mutation Rate
  • RNA, Guide, CRISPR-Cas Systems / metabolism
  • Ribonucleoproteins / metabolism*
  • Zea mays / cytology
  • Zea mays / genetics*

Substances

  • DNA, Plant
  • Multiprotein Complexes
  • RNA, Guide, CRISPR-Cas Systems
  • Ribonucleoproteins