Targeted mutagenesis in rice using CRISPR-Cas system  

作　　者：Jin Miao Dongshu Guo Jinzhe Zhang Qingpei Huang Genji Qin Xin Zhang Jianmin Wan Hongya Gu Li-Jia Qu 

机构地区：[1]State Key Laboratory for Protein and Plant Gene Research, Peking-Tsin- ghua Center for Life Sciences, College of Life Sciences, Peking University, Beijing 100871, China [2]National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China [3]The National Plant Gene

出　　处：《Cell Research》2013年第10期1233-1236,共4页细胞研究（英文版）

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.

关 键 词：CAS系统 定向诱变 模式生物基因组 水稻 DNA双链断裂 非同源末端连接 基因功能分析 重复序列 

分 类 号：Q987.1[生物学—遗传学] TU243[生物学—人类学]