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Targeted genome modification of crop plants using a CRISPR-Cas system
Qiwei Shan,Yanpeng Wang,Jun Li,Yi Zhang,Kunling Chen,Zhen Liang,Kang Zhang,Jinxing Liu,Jianzhong Jeff Xi,Jin-Long Qiu & Caixia Gao
Nature Biotechnology
DOI:10.1038/nbt.2650
Abstract
We describe the use of the cluster regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) system (CRISPR/Cas) system involving RNA-guided endonucleases to introduce specific mutations in rice and wheat. For four rice genes and one wheat gene, the frequencies of sgRNA:Cas9 induced mutations were 14.5 to 38.0% in protoplasts, and 4.0 to 9.4% in regenerated rice plants. We also demonstrate homology-directed targeted gene modification in rice using a single-stranded DNA (ssDNA) oligo as a donor template. Our findings establish that the sgRNA:Cas9 system can be used for rice and wheat genome modification, the first plants shown to be amenable to this gene editing approach.
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