本节将重点介绍这样一种策略- - FLEX开关,它利用重组关闭一个基因的表达,同时开启另一个基因的表达。 什么是FLEX开关 FLEX (或"翻转-切除")开关被设计为一种基因工具,研究人员利用位点特异性重组在体内有条件地操纵基因表达。FLEX开关利用了位点特异性重组酶(SSRs) Cre和FLP的方向特异性。SSRs在靶位点结合DNA,诱...
1. ConVERGD系统的开发与验证:研究团队首先成功开发了ConVERGD系统,该系统融合了自切割核糖核酸酶和FLEx开关,能够在重组酶(如Cre和Flp)的作用下表达特定的分子载荷。通过体外细胞实验,验证了ConVERGD系统在不同条件下表达的特异性。 2. ConVERGD系统的扩展:为了增强系统的多特征交叉靶向能力,研究团队对ConVERGD系统进...
AAV phSyn1-FSF-FLEX-ChR2(H134R)-EYFP-WPRE-bGHpA catalog : 65454 more info or order: Addgene product webpage citations: 1 Reference Madisen L, Garner A, Shimaoka D, Chuong A, Klapoetke N, Li L,et al. Transgenic mice for intersectional targeting of neural sensors and effectors with...
Data points represent independent transfections with the following constructs: 56 (FLEx(FRT)eGFP alone or +pDIRE), 6 (CV-eGFP-W3SL +Flp, +pDIRE; CV-eGFP-WPRE, +Flp, +pDire; Ef1a-CV-eGFP-WPRE, +pDIRE) 5 (INTRSECT-EYFP, +Cre, +Flp, +pDIRE; CV-eGFP-W3SL, +Cre; CV-eGFP-...
此外,袋具有带C-Flex管和Luer或MPC连接器的自由端口,用于添加培养基、接种物和碱。将带有Luer隔膜的浸入管用于取样。 对于灌流实验,使用带有集成1.2μm灌流膜的特殊的袋。将膜固定在袋的底部,形成用于去除无细胞培养基的区室。将膜的表面用由此产生的波的每一次摇动冲洗,其确保低污染和长培养时间。使用 B控制...
Quantitative real-time PCR was performed using PowerUp SYBR Green Master Mix in QuantStudio™ 6 7 Flex Real-Time PCR System (Life Technologies, Thermo Fisher Scientific). Human and murine SGPL1 mRNA and levels of cytokines were measured. Cq values of samples were normalized to the ...
(A) Left, AAV1-hSyn-Cre was injected into V1 of Ai14 mice, followed by a second injection of AAV1-CAG-FLEX-GFP into SC. Middle, GFP-labeled neurons in SC-sg were also tdTomato+. Right four panels, GFP-labeled axons in various regions (LP, pretectal area, PBG, and cuneiform nu...
pAAV-CAG-FLEX-EGFP-WPRE 荧光蛋白 CAG EGFP Cre依赖表达 pAAV-CAG-EGFP-3xFLAG-WPRE 荧光蛋白 CAG EGFP 直接表达 pAAV-CAG-mCherry-3xFLAG-WPRE 荧光蛋白 CAG mCherry 直接表达 pAAV-CAG-EGFP-P2A-3xFLAG-WPRE 荧光蛋白 CAG EGFP 直接表达 pAAV-CAG-mCherry-P2A-3xFLAG-WPRE 荧光蛋白 CAG mCherry ...
1. ConVERGD系统的开发与验证:研究团队首先成功开发了ConVERGD系统,该系统融合了自切割核糖核酸酶和FLEx开关,能够在重组酶(如Cre和Flp)的作用下表达特定的分子载荷。通过体外细胞实验,验证了ConVERGD系统在不同条件下表达的特异性。 2. ConVERGD系统的扩展:为了增强系统的多特征交叉靶向能力,研究团队对ConVERGD系统进...
AAV2/1-hSyn-Flp、 AAV2/9-Cre/Flp-NpHR-EYFP、AAV2/9-Cre/Flp-ChR2-EYFP、AAV2/9-Cre/Flp-EYFP 、AAV2/9-Cre/Flp-EYFP、 AAV2/9-Cre/Flp-EYFP 、Retro-AAV2/2-CAG-Flex-Flp 、AAV2/9-fDIO-GCaMP6s 、AAV2/9-DIO-taCasp3-TEVp、AAV2/9-hSyn-Cre、AAV2/9-DIO-taCasp3-TEVp、AAV2/...