产品简介
YINGGUANGSU(Luciferase)SHIZIRANJIEZHONGNENGGOUCHANSHENGSHENGWUFAGUANGDEMEIDETONGCHENG,QIZHONGZUIYOUDAIBIAOXINGDESHILAIZIYINGHUOCHONGTINEI(Firefly)HEHAISHEN(Renilla)TINEIDELIANGLEIYINGGUANGSUMEI,FENBIEMINGMINGWEIF-LuciferaseHER-Luciferase。ZAIYINGGUANGSUMEIDECUIHUAXIA,YINGGUANGSU DIWUKEYIGAOXIAODEZHUANBIANCHENGYANGYINGGUANGSU,TONGSHIFACHUQIANGLIEDEGUANG。
JIANYUCI,YANJIUZHELIYONGYINGGUANGSUMEIKAIFACHUYITAOJIQILINGMINQIESHIYONGFANGBIANDEJIYINBAOGAOXITONG。MUQIANZHEIYIXITONGBEIGUANGFANYONGYUZHUANLUYINZIJIEHEWEIDIANYUQIDONGZIHUOXINGFENXI、RNAJIANGJIE、XINHAOZHUANDAO、YAOWUSHAIXUAN、DONGWUHUOTICHENGXIANGDENGLINGYU。
两种常用载体类型极其应用场景
pGL3-Basic---YONGYUZHUANLUYINZIJIEHEWEIDIANYUQIDONGZIHUOXINGFENXI。BADAIFENXIQIDONGZIQUDUANGOUJIANDAOF-LuciferaseSHANGYOU,HEZHUANLUYINZIGONGZHUANRANFENXIF-LuciferaseHUOXINGJIKEFANYINGQIDONGZIDEHUOXINGGAODI。GAIZHILITONGCHANGXUYAOJIEHECHIXUXINGBIAODAR-LuciferaseDEZAITIZUOWEINEICANYIXIAOZHENGBUTONGYANGPINZHIJIANZHUANRANDEZHUANRANXIAOLV(CANKAOANLIYI)。
psiCHECK2---miRNAYUQIBADIANXIANGGUANFENXI,BAOKUOmiRNABAJIYINYANZHENG(CANKAOANLIERAHEB)、lncRNA (CANKAOANLISAN)YUcircRNA (CANKAOANLISI)XIFUmiRNAYANZHENGDENG。XUYAOBAmiRNAQIANZAIJIEHEBADIANKELONGDAOR-Luciferase (hRLuc) 3’UTRQUYU,RANHOUYUDAICEmiRNAGONGTONGZHUANRANXIBAONEICEDINGR-LuciferaseHUOXINGGAODI,F-Luciferase (hLuc+)ZUOWEIXIAOZHENGNEICANXIAOZHENGBUTONGYANGPINZHIJIANZHUANRANDEZHUANRANXIAOLV。
技术路线
经典应用案例一
LncRNAZHUANLUYINZIJIEHEWEIDIANYUQIDONGZIHUOXINGFENXI
LUNWENLAIYUAN: Wang, P., et al. (2014). "The STAT3-binding long noncoding RNA lnc-DC controls human dendritic cell differentiation." Science 344(6181): 310-313.
BENWENSHICAOXUETAOSHIYANSHIZAI2014NIANFABIAOZAIScienceSHANGYIPIANYANJIUXINGLUNWEN。ZUOZHETONGGUOXINPIANJIANDINGCHUDC (SHUTUZHUANGXIBAO)FAYUXIANGGUANDEYIGElncRNA,MINGMINGWEIlnc-DC。TONGSHIGAOTONGLIANGCEXUYEFAXIANLEZHEITIAOCHAYIlncRNADECUNZAI。JINERZUOZHETONGGUONorthern BlotYANZHENGLEZHEITIAOlnc-DC。YOUYUlnc-DCJIQITEYIHEWENDINGDIGAOBIAODAZAIDCFAYUGUOCHENGDEMOUYISHIQI,SUOYIZUOZHEJIASHElnc-DCKENENGSHIHEBIAOGUANYICHUANXUEDIAOKONGSHIJIANXIANGGUANDE。WEILEYANZHENGZHEIYIJIASHE,ZUOZHETONGGUORANSETIMIANYICHENDIANJIEHECEXUDEJISHU-CHIP-seqHEqPCRFAXIANlnc-DCDEZHUANLUQISHIWEIDIAN(TSS)FUJINFUJILEPol II、H3K4me3HEH3K27acDENGDANBAI(XIATUA),TISHIDCFAYUGUOCHENGZHONGBIAOGUANYICHUANXUEBIANHUAYUlnc-DCDEZHUANLUZENGQIANGSHIJIANXIANGDUIYING。JINERZUOZHETONGGUOXULIEFENXIFAXIANZAIlnc-DCDEZHUANLUQISHIWEIDIANFUJIN(+44-+50)CUNZAIYIGEJINGDIANDEPU.1JIEHEWEIDIAN。ZUOZHEJIASHEPU.1CANYULElnc-DCDETEYIXINGBIAODA。YUSHIZUOZHESHEJILERUXIASHIYANJINXINGLEJINYIBUDEYANZHENG(XIATUB)。
ZUOZHEBAlnc-DCDEQIDONGZIQUDUAN(-1447-+223)JIQIBUTONGDETUBIANTIBANBENFENBIEKELONGDAOF-LuciferaseBIAODAKUANGSHANGYOU,RANHOUHEZHUANLUYINZIPU.1BIAODAZHILIHUOZHEKONGZAITI(Vector)GONGTONGZHUANRANDAOMOSHIXIBAOXIHEK-293T,TONGGUOCELIANGYINGGUANGQIANGDUDEBIANHUAJIANJIEFANYINGZHUANLUHUOXINGDEGAODI。ZUIZHONGZHENGMINGZHUANLUYINZIPU.1TONGGUOJINGDIANJIEHEWEIDIAN(+44-+50)JIEDAOLElnc-DCZAIDCFAYUGUOCHENGZHONGDETEYIBIAODA。
经典应用案例二
LncRNATONGGUOXIFUmiRNADIAOKONGBAJIYINmRNADEBIAODA (ceRNA)
LUNWENLAIYUAN: Cesana, M., et al. (2011). "A long noncoding RNA controls muscle differentiation by functioning as a competing endogenous RNA." Cell 147(2): 358-369.
ZUOZHETONGGUOBIAODAFENXIFAXIANLEYITIAOJIROUFAYUXIANGGUANDElncRNA---lnc-MD1.QIANQIGONGNENGYANJIUFAXIANlnc-MD1KEYIDIAOKONGJIROUFENHUADESHIJIAN。XINXIXUEFENXIFAXIANZHEITIAOJIROUTEYIDElncRNASHANGBAOHAN36GEmiRNADEJIEHEWEIDIAN。RUGUOZHIKAOLVJIROUFAYUXIANGGUANDEmiRNAHEJIYIN,36GEHOUXUANmiRNALIMIANZHISHENGXIAmiR-133HEmiR-135。ZUOZHETONGGUOYINGGUANGSUMEISHIYANJINXINGLEYANZHENG。SHOUXIANZUOZHEBAYESHENGXINGJIQIQUESHImiR-133HEmiR-135JIEHEWEIDIANDELnc-DC1KELONGDAOJINR-LucDE3’UTR,RANHOUFENBIEYUBIAODAmiR-133HEmiR-135QIANTIDEZHILIGONGTONGZHUANRANDAOXIBAOJINXINGYINGGUANGSUMEIHUOXINGJIANCE(XIATUAHEB)。ZHENGMINGlnc-MD1QUESHICUNZAImiR-133HEmiR-135DEJIEHEWEIDIAN。
JINYIBUDEXINXIXUEYUCE,miR-133HEmiR-135FENBIEBAXIANGLIANGGEJIYIN- MAML1 HE MEF2C。TONGYANG,ZUOZHEHAISHITONGGUOYINGGUANGSUMEISHIYANJINXINGYANZHENG。YANJIUZHEBAMAML1 HE MEF2CDE3’UTR (WT)HEmiRNAJIEHEWEIDIANQUESHITUBIANTI(-mut)FENBIEKELONGZAIRLucBIAODAKUANGDEXIAYOU,RANHOUHEmiRNAGUOBIAODAZAITIGONGZHUANRANXIBAOJINXINGMEIHUOCEDING。JIEGUOXIANSHImiR-133HEmiR-135QUESHIFENBIEBAXIANGLEMAML1 HE MEF2CLIANGGEJIYIN(XIATUC)。ZUIHOUDEMOXINGSHIlnc-MD1TONGGUOceRNAJIZHIXIFUmiR-133HE135,ZHENGXIANGDIAOKONGLEMAML1 HE MEF2CDEBIAODA,CANYUDIAOKONGJIROUFAYU。
经典应用案例三
LncRNATONGGUOXIFUmiRNADIAOKONGQISHENGLIGONGNENG (ceRNA)
LUNWENLAIYUAN: Kallen, A. N., et al. (2013). "The imprinted H19 lncRNA antagonizes let-7 microRNAs." Molecular Cell 52(1): 101-112.
H19SHUYUFEICHANGBAOSHOUDEYINJIJIYINJIAZU,ZAIPEITAIFAYUHESHENGZHANGFANGMIANFAHUIJIQIZHONGYAODEGONGNENG。WEILEJINYIBUYANJIUlncRNA H19DEGONGNENG,ZUOZHETONGGUOXINXIXUEFENXIFAXIANH19BAOHANlet-7DE4GEJIEHEWEIDIAN。YUSHIZUOZHETUICEH19KENENGHUIYINGXIANGlet-7FAHUIZUOYONG。
WEILEYANZHENGZHEIYIYUCE,YANJIUZHEBA4GElet-7QIANZAIJIEHEWEIDIANCHUANLIANQILAIKELONGDAORLucDE3’UTRQU。RANHOUXIANHOUGENlet-7DEYIZHIXINGZAITI(XIATUAHEB)HEGUOBIAODAZAITI(XIATUC)FENBIEGONGZHUANRANDAOXIBAOXIJIANCEYINGGUANGSUMEIDEHUOXING,ZHENGMINGLet-7DEJIEHEWEIDIANQUESHICUNZAIQIEFAHUIGONGNENG。JINYIBUZHENGMINGH19TONGGUORNA spongeDEJIZHIYINGXIANGlet-7JIAZUBAJIYINDEBIAODACANYUJIROUFAYUDIAOKONGGUOCHENG。
经典应用案例四
circRNATONGGUOXIFUmiRNADIAOKONGQIBAJIYINmRNADEBIAODA (ceRNA)
LUNWENLAIYUAN: Zheng, Q., et al. (2016). "Circular RNA profiling reveals an abundant circHIPK3 that regulates cell growth by sponging multiple miRNAs." Nat Commun 7: 11215.
HUANZHUANGRNA (circRNA)SHIFEIBIANMARNALINGYUYILEIXINDEMINGXINGFENZI,YANJIUZHEDUIQIGONGNENGHAIZHIZHISHENSHAO。
BENWENZHONG,ZUOZHEJIANDINGCHUYIGEBIAODALIANGJIAOGAODEHUANZHUANGRNAFENZI-circHIPK3. circHIPK3LAIYUANYUJIYINHIPK3DIERGEWAIXIANZI。RNAiGANRAOcircHIPK3KEYIYINGXIANGXIBAODESHENGZHANG。WEILEJINYIBUYANJIUQIJIZHI,ZUOZHETONGGUOXULIEFENXIFAXIANcircHIPK3HANYOUYIXIELIEmiRNAJIEHEWEIDIAN。WEILECONGZHONGSHAIXUANCHUZHENZHENGDEmiRNA,ZUOZHEGOUJIANLEJIYUYINGGUANGSUMEIDESHAIXUANXITONG---BAcircHIPK3GOUJIANDAOLuciferaseDE3’UTRQUYU,RANHOUFENBIEYU424GEmiRNA mimicsGONGZHUANRANDAOMOSHIXIBAOXIHEK-293TNEIJINXINGSHAIXUANYANZHENG,ZUIZHONGZUOZHESHAIXUANCHU9GEHOUXUANDEmiRNA。TONGGUOGONGNENGSHIYANHEGONGBIAODAFENXIJINYIBUYANZHENGDEDAOmir-124SHIYANGXINGHOUXUANmiRNA。ZUIZHONGZUOZHEZHENGMINGcircHIPK3TONGGUOXIFUmiR-124ZHENGXIANGDIAOKONGLEQIBAJIYINIL6R HE DLX2DEBIAODA,YINGXIANGLEXIBAODESHENGZHANG。
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