Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease

Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease To the Editor: Sustainable intensification of crop production is essential to ensure food demand is matched by supply as the human population continues to increase . This will require high-yielding crop varieties that can be grown sustainably with fewer inputs on less land. Both plant breeding and genetic modification (GM) methods make valuable contributions to varietal improvement, but targeted genome engineering promises to be critical to elevating future yields. Most such methods require targeting DNA breaks to defined locations followed by either nonhomologous end joining (NHEJ) or homologous recombination . Zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) can be engineered to create such breaks, but these systems require two different DNA binding proteins flanking a sequence of interest, each with a C-terminal FokI nuclease module. We report here that the bacterial clustered, regularly interspaced, short palindromic repeats (CRISPR) system, comprising a CRISPR-associated (Cas)9 protein and an engineered single guide RNA (sgRNA) that specifies a targeted nucleic acid sequence , is applicable to plants to induce mutations at defined loci. To test the potential of the Cas9 system to induce gene knockouts in plants, we took advantage of Agrobacterium tumefaciens –mediated transient expression assays (agroinfiltration) to http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nature Biotechnology Nature Publishing Group (NPG)

Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease

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Publisher
Nature Publishing Group (NPG)
Copyright
Copyright © 2013 Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.
ISSN
1087-0156
eISSN
1546-1696
D.O.I.
10.1038/nbt.2655
Publisher site
See Article on Publisher Site

Abstract

To the Editor: Sustainable intensification of crop production is essential to ensure food demand is matched by supply as the human population continues to increase . This will require high-yielding crop varieties that can be grown sustainably with fewer inputs on less land. Both plant breeding and genetic modification (GM) methods make valuable contributions to varietal improvement, but targeted genome engineering promises to be critical to elevating future yields. Most such methods require targeting DNA breaks to defined locations followed by either nonhomologous end joining (NHEJ) or homologous recombination . Zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) can be engineered to create such breaks, but these systems require two different DNA binding proteins flanking a sequence of interest, each with a C-terminal FokI nuclease module. We report here that the bacterial clustered, regularly interspaced, short palindromic repeats (CRISPR) system, comprising a CRISPR-associated (Cas)9 protein and an engineered single guide RNA (sgRNA) that specifies a targeted nucleic acid sequence , is applicable to plants to induce mutations at defined loci. To test the potential of the Cas9 system to induce gene knockouts in plants, we took advantage of Agrobacterium tumefaciens –mediated transient expression assays (agroinfiltration) to

Journal

Nature BiotechnologyNature Publishing Group (NPG)

Published: Aug 8, 2013

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