gaotongliangsgrnawenkushaixuanfuwushiyongquanjiyinzuhuotedingtongludesgrnawenkuzaizhidingxibaoneitongguogongnengxingshaixuan、fujiyijisuihoudepcrkuozengheshenducexufenxi，wajueyubiaoxingxiangguandejiyinhuozaiquanjiyinzufanweineishaixuanyaowuxinbadian。sgrnawenkujiehegaotongliangshaixuanjishuzaiyaowuyanfa、jiyingongnengyanjiu、fenzizhenduan、jibingzhiliaohezuowuyuzhongdenglingyujunfahuidezhongyaodezuoyong。anbiqishengwudecrispr/cas9pingtaiketigonggaotongliangsgrnawenkushaixuanfuwu，womendefuwuhangaisgrnawenkugoujian，xibaozhuanran，xibaoshaixuan，gaotongliangcexu，gongnengjiyinshaixuanhegongnengjizhifenxi。womenbujinnengbangzhuguangdakehuzhibeiquanjiyinzuhuotedingtongludesgrnawenku，haikeyiwanchengxiayoudexibaoshaixuanhegaotongliangcexu，wajuexiangguandegongnengjiyinhuoyaowubadianbingjiexiqijizhi。

sgRNA文库筛选服务流程

1. sgRNA设计与文库构建：gaotongliangdesgrnawenku。

2. 细胞转染与筛选：目的细胞侵染和功能实验；阳性细胞筛选。

3. 功能基因筛选：PCR扩增sgRNA；高通量测序及数据分析；单基因功能验证。

4. 功能基因机制研究：xibaoshengwuxuegongnengshiyan；biaodapuxinpianfenxitonglujizhi；xiayoujiyingongnengyanjiu。

anbiqishengwuweiguangdakehutigongsgrnawenkushaixuanfuwu，baokuocrisprqiaochushaixuan，crisprjihuoshaixuanhecrispryizhishaixuan。

• CRISPR敲除筛选

cirsprjiyinqiaochushaixuankeyiwajuejiyinzufanweineidegongnengqueshixingjiyin，haiyouzhuyujieshiyizhijiyindexingongneng，jiehebutongdehuahewushaixuankeyiyouxiaodifaxianyaowudexinbadian。

• CRISPR激活筛选

crisprjihuoshaixuandeyuanlishijiangqiegehuoxingqueshidedcas9yuzhuanlujihuoyinzi（ruvpr，vp64）ronghe，zaisgrnadeyindaoxiabaxiangjiyinshangyouzhuanlujihuoquyubingyouxiaojihuojiyindebiaoda。crisprjihuoshaixuanyijingbeiyingyongyugaotongliangdegongnenghuodexingjiyinshaixuan，haiyouzhuyuyanjiuzaitianrantiaojianxiabiaodaliangjiaodidejiyin。

• CRISPR抑制筛选

CRISPR抑制筛选的原理是将切割活性缺失的dCas9与转录抑制因子（如：KRAB）融合，在sgRNA的引导下有效的抑制靶基因的表达。与敲除筛选相比，CRISPR抑制筛选是通过下调基因表达来筛选功能基因，是揭示敲除致死型基因功能的有效方法。

高通量sgRNA文库筛选应用

• tansuoyaowuzuoyongjizhi：yaowubadianquedingyuyanzheng

• xunzhaozhongliuzhiliaobadian：shangxiayoudiaokongjizhifenxi

• tanjiudaixiejibingzhiliaojizhi：daixietongludiaojiejizhifenxi

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