title: Genome editing in maize directed by CRISPR-Cas9 ribonucleoprotein complexes.
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.
authors.affiliation: Trait Enabling Technologies, DuPont Pioneer, Johnston, Iowa 50131, USA.
authors.forename: Sergei
authors.initials: S
authors.lastname: Svitashev
authors.affiliation: Trait Enabling Technologies, DuPont Pioneer, Johnston, Iowa 50131, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Schwartz
authors.affiliation: Trait Enabling Technologies, DuPont Pioneer, Johnston, Iowa 50131, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Lenderts
authors.affiliation: Trait Enabling Technologies, DuPont Pioneer, Johnston, Iowa 50131, USA.
authors.forename: Joshua K
authors.initials: JK
authors.lastname: Young
authors.affiliation: Trait Enabling Technologies, DuPont Pioneer, Johnston, Iowa 50131, USA.
authors.forename: A
authors.initials: A
authors.lastname: Mark Cigan
mesh_terms: D000483:Alleles; D001483:Base Sequence; D019470:Biolistics; D064113:CRISPR-Cas Systems / Q000235:genetics*; D018744:DNA, Plant / Q000378:metabolism; D000072669:Gene Editing; D018014:Gene Transfer Techniques; D017343:Genes, Plant; D046912:Multiprotein Complexes / Q000378:metabolism*; D016296:Mutagenesis / Q000235:genetics; D009154:Mutation / Q000235:genetics; D059645:Mutation Rate; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism*; D003313:Zea mays / Q000166:cytology / Q000235:genetics*
chemical_list: D018744:DNA, Plant; D046912:Multiprotein Complexes; D000094704:RNA, Guide, CRISPR-Cas Systems; D012261:Ribonucleoproteins