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Targeted mutagenesis in rice using CRISPR-Cas system期刊论文

作者: Jin Miao Dongshu Guo Jinzhe Zhang Qingpei Huang Genji Qin Xin Zhang Jianmin Wan Hongya Gu Li-Jia Qu
DOI:10.1038/cr.2013.123

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页码: 1233-1236
被引频次: 306
出版者: INST BIOCHEMISTRY & CELL BIOLOGY,Nature Publishing Group
ISSN: 1001-0602
卷期: Volume:23    Issue:10
语言: English
摘要: Genome editing of model organisms is essential for gene function analysis and is thus critical for human health and agricultural production. The current technolo- gies used for genome editing include ZFN (zinc-finger nuclease), meganucleases, TALEN (Transcription activa- tor-like effector nucleases), etc. [1]. These technologies can generate double stranded breaks (DSBs) to either disrupt gene function through generation of premature stop codons by non-homologous end joining (NHEJ) pathway, or to facilitate gene targeting through homolo- gous recombination (HR) with an incoming template. Recently, a new technology for genome editing, CRISPR (Clustered Regularly Interspaced Short Palindromic Re- peats)/Cas (CRISPR-associated) systems, has been de- veloped [2]. CRISPR/Cas systems are adaptive defense systems in prokaryotic organisms to fight against alien nucleic acids [3]. The spacer sequences acquired from foreign DNA are positioned between host repeats, and transcribed together as CRISPR RNA (crRNA). In the type II CRISPR system, a single nuclease Cas9, guided by a dual-crRNA:tracrRNA, is sufficient to cleave cog- nate DNA homologous to the spacer [2]. Efficient cleav- age also requires the presence of protospacer adjacent motif (PAM) 5'-NGG-3' following the spacer sequence. The dual-crRNA:tracrRNA has been further streamlined to a single RNA chimera, called sgRNA (single guide RNA) [2]. Compared with protein-guided technologies, CRISPR/Cas system is much easier to implement, as only short guide RNAs need to be customized to target the genes of interest. Up to now, the CRISPR/Cas sys- tem has been successfully applied to efficient genome editing in many eukaryotic organisms including human [1], mice [4], zebra fish [5], fly [6], worm [7], and yeast [8]. However, the application of CRISPR/Cas system in plants has not been reported. Rice (Oryza sativa L.) is a major staple crop in the grass family (Poaceae), feed- ing half of the world's population.
相关主题: CAS系统, DNA双链断裂, 重复序列, 模式生物基因组, 水稻, 非同源末端连接, 定向诱变, 基因功能分析, GENOME, CELL BIOLOGY, Mutagenesis, Oryza - genetics, Genes, Plant, CRISPR-Cas Systems, Letter to the Editor,

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