Multiplex and homologous recombination-mediated genome editing in Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9

Journal Article / 2013

Li, Jian-Feng; Norville, Julie E.; Aach, John; McCormack, Matthew; Zhang, Dandan; Bush, Jenifer; Church, George M.; Sheen, Jen

Elucidation and manipulation of human, animal and plant genomes is key to basic biology research, medical advances and crop improvement. The development of targeted genome editing, particularly homologous recombination–based gene replacement, is of great value in all organisms. Recent advances in engineered nucleases with programmable DNA-binding specificities, such as zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), have provided valuable means to create targeted mutations in metazoan and plant genomes with high specificity 1–6 . However, these technologies demand elaborate design and assembly of individual DNA-binding proteins for each DNA target site 1–6 . Recently, a simple, versatile and efficient genome engineering

amplicon; Arabidopsis; Arabidopsis Proteins; Arabidopsis thaliana; bacterial growth; Base Sequence; Caspase 9; cell division; cellular apoptosis susceptibility protein DNA binding protein; DNA directed DNA polymerase gamma; double stranded DNA; escherichia coli; gene replacement therapy; genetic engineering; genetic recombination; Genome, Plant; genomic DNA; guide RNA; homologous recombination; human; letter; mutagenesis; Nicotiana benthamiana; onhuman; oxidation; Oxidoreductases; priority journal; procaspase 9; protein expression; Protein Kinases; protein localization; protoplast; RNA; sequence analysis; tobacco; zinc finger nuclease

Nature biotechnology

8

688–691

31

10.1038/nbt.2654

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