Dual fluorescent protein reporters for studying cell behaviors in vivo.
Dual fluorescent protein reporters for studying cell behaviors in vivo.
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DOI：
10.1002/dvg.20565
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发表时间：
2009-10
期刊：
影响因子：
1.5
通讯作者：
Behringer, Richard R.
中科院分区：
文献类型：
--
作者：
Stewart, M. David;Jang, Chuan-Wei;Hong, Nicholas W.;Austin, Anthony P.;Behringer, Richard R.
荧光蛋白(FP)是显示活细胞及其行为的有用工具。调节亚细胞定位的蛋白质结构域已与FP融合，以突出细胞结构。与组蛋白H2B融合的FPS结合到染色质中，从而使核事件可视化。FPS与糖基磷脂酰肌醇锚定信号序列(GPI)融合，标记质膜，突出细胞形状。因此，包含这两种类型的FP融合的报告基因将允许有效地监测细胞的形状、运动、有丝分裂阶段、凋亡和其他细胞活动。在这里，我们报告了一个二进制颜色编码系统，它使用四个不同颜色的荧光蛋白报告器，产生16个不同的颜色代码来标记培养中和转基因小鼠中活细胞的细胞核和质膜。作为该系统在体内的初步测试，人类泛素C(UBC)基因的启动子被用来广泛表达其中一个色码报告。该报告在胚胎中得到了广泛的表达；然而，雄性和雌性转基因小鼠都是不育的。相比之下，小鼠Oct4/Pou5f1基因的启动子连接到两个不同的颜色编码记者，这些小鼠是可生育的。荧光标记的原始生殖细胞的延时电影展示了色码系统在可视化细胞行为方面的应用。这套新的荧光蛋白报告器应该是标记不同的细胞群体和研究它们在体内复杂组织中行为的有用工具。
Fluorescent proteins (FPs) are useful tools for visualizing live cells and their behaviors. Protein domains that mediate subcellular localization have been fused to FPs to highlight cellular structures. FPs fused with histone H2B incorporate into chromatin allowing visualization of nuclear events. FPs fused to a glycosylphosphatidylinositol anchor signal sequence (GPI) label the plasma membrane, highlighting cellular shape. Thus, a reporter gene containing both types of FP fusions would allow for effective monitoring of cell shape, movement, mitotic stage, apoptosis and other cellular activities. Here we report a binary color-coding system using four different colored fluorescent protein reporters that generates 16 distinct color codes to label the nuclei and plasma membranes of live cells in culture and in transgenic mice. As an initial test of this system in vivo, the promoter of the human Ubiquitin C (UBC) gene was used to widely express one of the color code reporters. Widespread expression of the reporter was attained in embryos; however, both male and female transgenic mice were infertile. In contrast, the promoter of the mouse Oct4/Pou5f1 gene linked to two different color code reporters specifically labeled blastocysts, primordial germ cells, and postnatal germ cells and these mice were fertile. Time-lapse movies of fluorescently-labeled primordial germs cells demonstrate the utility of the color code system to visualize cell behaviors. This set of new fluorescent protein reporters should be a useful tool for labeling distinct cell populations and studying their behaviors in complex tissues in vivo.
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影响因子：
5.4
作者：
Trichas, Georgios;Begbie, Jo;Srinivas, Shankar
通讯作者：
Srinivas, Shankar
影响因子：
1.5
作者：
Hsiao, YC;Chang, HH;Tsai, TF
通讯作者：
Tsai, TF
影响因子：
2.7
作者：
PALMIERI, SL;PETER, W;SCHOLER, HR
通讯作者：
SCHOLER, HR
影响因子：
2.6
作者：
Li, X;Harrell, RA;Fraser, MJ
通讯作者：
Fraser, MJ
DOI：
10.1002/ar.20821
发表时间：
2009-03
影响因子：
2
作者：
Larina, Irina V.;Shen, Wei;Kelly, Olivia G.;Hadjantonakis, Anna-Katerina;Baron, Margaret H.;Dickinson, Mary E.
通讯作者：
Dickinson, Mary E.