Genome editing in maize directed by CRISPR–Cas9 ribonucleoprotein complexes

作者：Sergei Svitashev, Christine Schwartz, Brian Lenderts, Joshua K. Young, A. Mark Cigan · 发表于：Nature Communications · 年份：2016 · DOI：10.1038/ncomms13274 · 被引用次数：848 · 研究领域：CRISPR and Genetic Engineering、Insect symbiosis and bacterial influences、Plant Virus Research Studies

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.