DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins

Nat Biotechnol. 2015 Nov;33(11):1162-4. doi: 10.1038/nbt.3389. Epub 2015 Oct 19.

Abstract

Editing plant genomes without introducing foreign DNA into cells may alleviate regulatory concerns related to genetically modified plants. We transfected preassembled complexes of purified Cas9 protein and guide RNA into plant protoplasts of Arabidopsis thaliana, tobacco, lettuce and rice and achieved targeted mutagenesis in regenerated plants at frequencies of up to 46%. The targeted sites contained germline-transmissible small insertions or deletions that are indistinguishable from naturally occurring genetic variation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics
  • Arabidopsis / physiology
  • Base Sequence
  • CRISPR-Cas Systems / genetics*
  • Gene Knockout Techniques
  • Genetic Engineering / methods*
  • Genome, Plant / genetics*
  • Molecular Sequence Data
  • Oryza / genetics
  • Oryza / physiology
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Plants, Genetically Modified / genetics*
  • Plants, Genetically Modified / physiology
  • Protoplasts
  • Ribonucleoproteins / genetics*
  • Ribonucleoproteins / metabolism
  • Tobacco / genetics
  • Tobacco / physiology

Substances

  • Plant Proteins
  • Ribonucleoproteins