A simple and efficient system for regulating gene expression in human pluripotent stem cells and derivatives.
A simple and efficient system for regulating gene expression in human pluripotent stem cells and derivatives.
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DOI：
10.1002/stem.1653
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发表时间：
2014-05
期刊：
影响因子：
5.2
通讯作者：
Zhang, Su-Chun
中科院分区：
文献类型：
--
作者：
Qian, Kun;Huang, CindyTzu-Ling;Chen, Hong;Blackbourn, Lisle W.;Chen, Yuejun;Cao, Jingyuan;Yao, Lin;Sauvey, Cornall;Du, Zhongwei;Zhang, Su-Chun
人多能干细胞（hPSC）及其后代中可调控的转基因表达对于以时间和空间方式剖析基因功能通常是必需的。然而，由于在干细胞扩增和/或分化期间随机插入的基因的沉默，具有诱导型转基因表达的hPSC系，特别是在分化的后代中，尚未建立。在这里，我们报告了TALEN（转录激活因子样效应物核酸酶）介导的靶向AAVS 1位点以产生多功能条件性hPSC系的用途。hPSC及其衍生物中的转基因（GFP和功能基因）表达不仅持续，而且在体外和体内响应于多西环素而受到严格调控。我们修饰了供体构建体，使得任何感兴趣的基因可以容易地插入以产生具有条件性转基因表达的hPSC系。这项技术将大大改善我们研究人类干细胞的方式。
Regulatable transgene expression in human pluripotent stem cells (hPSCs) and their progenies is often necessary to dissect gene function in a temporal and spatial manner. However, hPSC lines with inducible transgene expression, especially in differentiated progenies, have not been established due to silencing of randomly inserted genes during stem cell expansion and/or differentiation. Here, we report the use of TALEN (transcription activator–like effector nucleases)-mediated targeting to AAVS1 site to generate versatile conditional hPSC lines. Transgene (both GFP and a functional gene) expression in hPSCs and their derivatives was not only sustained but also tightly regulated in response to doxycycline both in vitro and in vivo. We modified the donor construct so that any gene of interest can be readily inserted to produce hPSC lines with conditional transgene expression. This technology will substantially improve the way we study human stem cells.
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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
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Jaenisch, Rudolf
影响因子：
17.3
作者：
Gaj, Thomas;Gersbach, Charles A.;Barbas, Carlos F., III
通讯作者：
Barbas, Carlos F., III
影响因子：
5.3
作者：
Cherry, SR;Biniszkiewicz, D;Jaenisch, R
通讯作者：
Jaenisch, R
影响因子：
5.4
作者：
Ogata, T;Kozuka, T;Kanda, T
通讯作者：
Kanda, T
影响因子：
5.2
作者：
Xia, Xiaofeng;Ayala, Melvin;Zhang, Su-Chun
通讯作者：
Zhang, Su-Chun