GAPTrap: A Simple Expression System for Pluripotent Stem Cells and Their Derivatives.
GAPTrap: A Simple Expression System for Pluripotent Stem Cells and Their Derivatives.
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DOI：
10.1016/j.stemcr.2016.07.015
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发表时间：
2016-09-13
影响因子：
5.9
通讯作者：
Stanley, Edouard G.
中科院分区：
文献类型：
--
作者：
Kao, Tim;Labonne, Tanya;Niclis, Jonathan C.;Chaurasia, Ritu;Lokmic, Zerina;Qian, Elizabeth;Bruveris, Freya F.;Howden, Sara E.;Motazedian, Ali;Schiesser, Jacqueline V.;Costa, Magdaline;Sourris, Koula;Ng, Elizabeth;Anderson, David;Giudice, Antonietta;Farlie, Peter;Cheung, Michael;Lamande, Shireen R.;Penington, Anthony J.;Parish, Clare L.;Thomson, Lachlan H.;Rafii, Arash;Elliott, David A.;Elefanty, Andrew G.;Stanley, Edouard G.
可靠地表达荧光报告基因或其他感兴趣的基因的能力对于使用人多能干细胞（hPSC）作为研究细胞命运和基因功能的平台是重要的。我们描述了一个简单的表达系统，命名为GAPTrap（GT），其中报告基因，包括GFP，mCherry，mTagBFP 2，luc 2，Gluc和lacZ插入到hPSC的GAPDH基因座中。携带GT载体变异的独立克隆表达了非常一致的报告基因水平。分化实验表明，报告基因表达可靠地维持在造血细胞，心脏中胚层，定形内胚层，和腹侧中脑多巴胺能神经元。类似地，对衍生自GT-lacZ hPSC的畸胎瘤的分析显示，β-半乳糖苷酶表达维持在代表三个胚层的衍生物的细胞类型谱中。因此，GAPTrap载体代表了一种稳健且直接的标记系统，其能够对PSC及其分化的衍生物进行不可磨灭的标记。GAPTrap载体系统将转基因靶向广泛表达的GAPDH基因座将转基因靶向GAPDH基因座产生可靠的转基因表达在该基因座的转基因在分化的细胞中稳健表达GAPTrap靶向人PSC系的产生简单而有效。在这篇文章中Stanley，Elefanty及其同事描述了一种简单的载体系统，GAPTrap，用于在多能干细胞及其分化衍生物中组成型表达转基因。GAPTrap载体将转基因靶向GAPDH基因座，确保在多种细胞类型中稳健和可靠的表达，并为细胞标记和基因过表达实验提供理想的基础。
The ability to reliably express fluorescent reporters or other genes of interest is important for using human pluripotent stem cells (hPSCs) as a platform for investigating cell fates and gene function. We describe a simple expression system, designated GAPTrap (GT), in which reporter genes, including GFP, mCherry, mTagBFP2, luc2, Gluc, and lacZ are inserted into the GAPDH locus in hPSCs. Independent clones harboring variations of the GT vectors expressed remarkably consistent levels of the reporter gene. Differentiation experiments showed that reporter expression was reliably maintained in hematopoietic cells, cardiac mesoderm, definitive endoderm, and ventral midbrain dopaminergic neurons. Similarly, analysis of teratomas derived from GT-lacZ hPSCs showed that β-galactosidase expression was maintained in a spectrum of cell types representing derivatives of the three germ layers. Thus, the GAPTrap vectors represent a robust and straightforward tagging system that enables indelible labeling of PSCs and their differentiated derivatives. GAPTrap vector system targets transgenes to the ubiquitously expressed GAPDH locus Targeting transgenes to the GAPDH locus yields reliable transgene expression Transgenes at this locus are robustly expressed in differentiated cells Generation of GAPTrap targeted human PSC lines is simple and efficient In this article Stanley, Elefanty, and colleagues describe a simple vector system, GAPTrap, for constitutively expressing transgenes in pluripotent stem cells and their differentiated derivatives. GAPTrap vectors target transgenes to the GAPDH locus, ensuring robust and reliable expression in a wide variety of cell types, and providing an ideal base for cell tagging and gene overexpression experiments.
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影响因子：
64.8
作者：
Kriks, Sonja;Shim, Jae-Won;Piao, Jinghua;Ganat, Yosif M.;Wakeman, Dustin R.;Xie, Zhong;Carrillo-Reid, Luis;Auyeung, Gordon;Antonacci, Chris;Buch, Amanda;Yang, Lichuan;Beal, M. Flint;Surmeier, D. James;Kordower, Jeffrey H.;Tabar, Viviane;Studer, Lorenz
通讯作者：
Studer, Lorenz
影响因子：
46.9
作者：
Hockemeyer, Dirk;Soldner, Frank;Beard, Caroline;Gao, Qing;Mitalipova, Maisam;DeKelver, Russell C.;Katibah, George E.;Amora, Ranier;Boydston, Elizabeth A.;Zeitler, Bryan;Meng, Xiangdong;Miller, Jeffrey C.;Zhang, Lei;Rebar, Edward J.;Gregory, Philip D.;Urnov, Fyodor D.;Jaenisch, Rudolf
通讯作者：
Jaenisch, Rudolf
DOI：
10.1126/science.1262110
发表时间：
2015-05-08
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
GTEx Consortium
通讯作者：
GTEx Consortium
影响因子：
4
作者：
Mossman, AK;Sourris, K;Elefanty, AG
通讯作者：
Elefanty, AG
影响因子：
5.2
作者：
Somers, Aba;Jean, Jyh-Chang;Sommer, Cesar A.;Omari, Amel;Ford, Christopher C.;Mills, Jason A.;Ying, Lei;Sommer, Andreia Gianotti;Jean, Jenny M.;Smith, Brenden W.;Lafyatis, Robert;Demierre, Marie-France;Weiss, Daniel J.;French, Deborah L.;Gadue, Paul;Murphy, George J.;Mostoslavsky, Gustavo;Kotton, Darrell N.
通讯作者：
Kotton, Darrell N.