RNA-guided genome editing a la carte  

作　　者：Philippe Horvath Rodolphe Barrangou 

机构地区：[1]DuPont Nutrition and Health, Dang -Saint- Romain, 86220, France [2]DuPont Nutrition and Health, Madison, WI 53716, USA [3]New ad- dress: Department of Food, Bioprocessing and Nutrition Scienee, North Carolina State Uni- versit) Raleigh, NC 27695, USA

出　　处：《Cell Research》2013年第6期733-734,共2页细胞研究（英文版）

摘　　要：Two recent papers in Science il lustrate how the prokaryotic CRIS PRCas immune system machinery, which typically targets invasive genetic elements such as viruses and plasmids, can be converted into a sophisticated molecular tool for nextgeneration human genome edit ing. The versatile Cas9 RNAguided endonuclease can be readily repro grammed using customizable small RNAs for sequencespecific single or doublestranded DNA cleavage. Molecular biologists can now order restriction enzymes h la carte, to target any DNA template and generate cleav age at any chosen site precisely, without the agony of finding the right cutter, or the caveat to settle for the nearest ac ceptable site. Even better, the chef also offers the choice of doublestranded DNA blunt cleavage, or strandspecific nicking. Indeed, two recent milestone reports in Science [1, 2] describe the diversion of the CRISPRCas bacte rial adaptive immune system in novel genome editing applications in mam malian cells. In bacteria and archaea, CRISPR （clustered regularly interspaced short palindromic repeats） and CRISPRasso ciated proteins （Cas） naturally provide adaptive immunity against viruses and plasmids through the uptake and stock piling of small fragments （sequences of about 30 bp named ＂spacers＂） of the encountered genetic elements into CRISPR arrays, in between direct re peats [3]. The subsequent transcription ofa CRISPR array into a precursor RNA （precrRNA）, followed by a maturation step in which the precrRNA is diced into smaller CRISPR RNAs （crRNAs）,produces a cocktail of small interfering RNAs that are loaded onto the patrolling Casencoded machinery involving helicase and nuclease activities to specifically target and destroy nucleic acid showing sequence complementar ity to the spacers [4]. Although generic, this scheme shows marked idiosyncra sies when considering the numerous and highly diverse CRISPRCas sys tems that have been examined [5]. For instance, the thoroughly studied Type II systems require an additional, smal

关 键 词：小分子RNA 人类基因组 编辑点 非同源末端连接 核酸内切酶 DNA裂解 双链DNA 位点特异性 

分 类 号：Q52[生物学—生物化学] Q987