Heritable gene-targeting with gRNA/Cas9 in rats
Dear Editor,
The rat is a preferred animal model in many research applications,especially in physiological,behavioral and translational studies [1].Although the rat embryonic stem (ES) cells were successfully isolated [2,3],the rat ES cell-based gene-targeting approach has not been widely adopted due to the technical difficulties in manipulating these cells.The first knockout rat was generated via microinjection of zinc finger nuclease (ZFN) into the embryos [4].In addition to ZFN,new genomeediting tools such as transcription activator-like effector nuclease (TALEN) [5] and the clustered,regularly interspaced,short palindromic repeats (CRISPR)/CRISPRassociated protein (Cas) system [6] have offered a rapid and efficient means of genome modification in many species.Technologies of ZFN and TALEN have made genetargeting in the rat genome more convenient and practical [7-9].The CRISPR/Cas system is the most recently developed technology for targeted genome modification in mammalian cells,bacteria,zebrafish and mice [10-12].This system requires a locus-specific CRISPR RNA (crRNA),a transactivating crRNA (tracrRNA) and a nuclease Cas9.Previous studies have shown that a chimeric RNA,consisting of a crRNA that recognizes the target sequence and a tracrRNA that recruites Cas9,could fulfill the combined functions of crRNA and tracrRNA [11,12].We previously reported highly efficient genome editing with the chimeric RNA-guided Cas9 system in mammalian cells and zebrafish [10].Here,we successfully extended this simple and efficient gRNA/Cas9 system to modify the rat genome.
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grants from the National Basic Research Program of China2012CB944501,2013CB531200 and 20IOCB529503;the Natural Science Foundation of China31271549 and 31221002
2013-11-29（万方平台首次上网日期，不代表论文的发表时间）
共4页
1322-1325
