一文掌握gRNA设计,常用方法汇总解析

CRISPRSHIZHIYOUXIAOBIANJIYOUJITINEIBUFENJIYINDEJISHU。MUQIANGENJUBUTONGDECasHESUANMEILAIFENLEIYIGONGYOUSANZHONGBUTONGDEXITONG。QIZHONGERXINGDEBIJIAOJIANDAN,SHIYICas9HESUANMEIYIJIXIANGDAORNA(gRNA)WEIHEXINZUCHENG。

如果要使用CRISPR/Cas9系统对基因组进行编辑,第一步也是最重要的一步,就是要设计gRNA。下面这篇文章就为大家汇总一下gRNA设计常用的方法:

 

 

一、首先着重介绍几个在线设计gRNA的工具

 

 

 ZHEZHONGTUIJIANLei Stanley Qi LabDE

 

ZHEIGEZAIXIANZHANDIANGONGNENGFEICHANGFENGFU,KEYIXUANZEJIYINBIANJIGONGJUDEQITAYONGTU(JIHUO,YIZHIJIYINBIAODADENG)(TU1-1)。XIAYIBUJIUSHIXUANZEWUZHONG(TU1-2)。DANZHIYOUCHANGJIANDE9ZHONG。ZAIXIAYIBUJIUSHIZHIJIESHURUJIYINDEMINGCHENG(HUOXULIE)JIKE(TU1-3)。

 

TU1-1 XUANZEBIANJILEIXING

 

 

TU1-2 XUANZEWUZHONG

 

           TU1-3 XUANZEJIYINMINGCHENG

                                                                                                                                                        

 

DANSHICHULAIDEsgRNAWEIZHIYOUYIXIEBINGBUZONGZAIATGDEXIAYOU(TU2SUOSHI,1,2,6,9,12,16,22JITIAOsgRNAWEIYUATGDESHANGYOU)。SUOYIGENJUWEIZHI,DUIYUZUOJIYINDEQIAOCHU(KO)ERYAN,QIANGLIEJIANYIXUANZEATGXIAYOUTONGCHANG100 aaYINEIFANWEINEIDEsgRNALAIYONG(BIRUTU3ZHONGDE3,5,7,10DENGJIAOXIAYOUDE);ERQIE,YIDINGYAOWEIYUCCDSQUYUNEI(CCDSSHIconsensus CDS,JIGONGGONGDECDSQUYU,SHIZHENDUIHENDUOGEZHUANLUBEN[isoforms]DINGYI,TU2ZHONGLUSETIAOKUANGJISHI。ZHEIGEHENZHONGYAO!!!)。

 

TU2

 

Feng Zhang lab:。 ZHEISHIZUIZAODEYIGESHEJIZHANDIAN。BAOKUOCHANGJIANDE16GEWUZHONG。XUANZHONGWUZHONG,BANIYAOSHEJIDEQUDUANXULIERENGDAOXIAMIANDEKUANGLIYUNXINGJIUXING。。DANQIANTISHINIZIJIYIJINGXUANHAOLEWEIZHI(TU3)

 

TU3

 

 

HAIYOUQITA,RUomictoolsXIAMIANDECas-Designer

ZHEIGEZHANDIANNEIRONGGENGJIAFENGFU,BIRUWUZHONGFUGAISHIZUIDUODE,BAOKUOZHIWU,KUNCHONGDENGDENG(TU4ZUO)。ZHEIGEZHANDIANYESHISHURUMUDEJIYINDEXULIEDINGZHIDE,BUTONGDESHIKEYIYOUJIAODADEZIYOUDUXUANZEWEIZHIYUCE。

 

 

TU4

 

 

HAIYOUYIXIESHANGYEYABOappGUANWANGDE,BIRUThermoXIAMIANDE(TU5):

WUZHONGJIAOSHAO,SHANGYEXINGMUDETEBIEQIANG,LEJIEXIAJIKE。