Cas9单克隆敲除细胞株

 

CRISPR/Cas XITONGQUANCHENGWEICHANGJIANHUIWENZHONGFUXULIECONGJI/CHANGJIANHUIWENZHONGFUXULIECONGJIGUANLIANDANBAIXITONG(clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins)。ZHEIYIXITONGSHIYUANYUXIJUNZHONGDEYIZHONGHOUTIANMIANYIXITONG(TU1)。GAIXITONGKEYIBAQINRUXIJUNDESHIJUNTIYICHUANWUZHIZHENGHEDAOZISHENJIYINZUYIGEHUOZHEDUOGECRISPRWEIDIANZUOWEIYONGJIUDE“JIYI”,DANGXIJUNBEIZAICIRUQINDESHIHOU,CRISPR WEIDIANBEIZHUANLUSHENGCHENG CRISPR RNAs(crRNAs)。crRNA SUIHOUHUIYINDAO DNAJIANQIEMEI Cas9 GENJUXULIEHUBUDEYUANZEJIANQIERUQINDEWAIYUANHESUANXULIE,XIAOMIEWAILAIYICHUANWUZHI,FAHUIBAOHUGONGNENG(TU2)。JIANERYANZHI,YUANYU CRISPR/Cas XITONGDEJIYINZUBIANJIJISHUBEIGONGCHENGHUAWEIYIDUANJIEDAODINGWEIZUOYONGDE RNA XULIE(guide RNAHEcrRNAHEERWEIYIDEZAHERNAXULIE,CHENGWEIsgRNA)JIASHANGYIGEDNASHUIJIEMEICas9ZUCHENGDEERYUANXITONG。


  研究者把这一系统工程化成可以用于编辑哺乳动物细胞的CRISPR/Cas9系统。具体包括适于哺乳动物表达的 Cas9 基因和定位作用的 sgRNA(图3)。方便起见,研究者把这两者放在了一个载体之中(all in one plasmid)。如果计划利用这一系统编辑某个目的基因,我们唯一要做的就是定制一条有效的sgRNA。

 


 

TU1  CRISPR/Cas XITONGLAIYUANYUXIJUNDE“MIANYIXITONG”

 

 

 

TU2 XIJUNLIYONG CRISPR/Cas XITONGGONGJISHIJUNTIRUQINSHIYITU