Live visualization of genomic loci with BiFC-TALE.
Live visualization of genomic loci with BiFC-TALE.
复制标题
使用 BiFC-TALE 实时可视化基因组位点。
DOI：
10.1038/srep40192
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发表时间：
2017-01-11
影响因子：
4.6
通讯作者：
Tang C
中科院分区：
文献类型：
--
作者：
Hu H;Zhang H;Wang S;Ding M;An H;Hou Y;Yang X;Wei W;Sun Y;Tang C
跟踪基因组位点的动态对于理解基本细胞内过程的机制是重要的。然而，这些基因座在活细胞中的荧光标记和成像一直具有挑战性。其中一个主要原因是活细胞核中弥漫性全长荧光蛋白（FPs）的背景荧光导致图像的低信本比（SBR），阻碍了活细胞基因组标记方法的应用。结合双分子荧光互补（BiFC）和转录激活因子样效应（TALE）技术，我们开发了一种标记基因组位点的新方法（BiFC-TALE），该方法大大降低了背景荧光水平。使用bbc - tale，与使用全长FP的方法相比，我们通过成像活细胞中的端粒和着丝粒证明了显著改善的SBR。
Tracking the dynamics of genomic loci is important for understanding the mechanisms of fundamental intracellular processes. However, fluorescent labeling and imaging of such loci in live cells have been challenging. One of the major reasons is the low signal-to-background ratio (SBR) of images mainly caused by the background fluorescence from diffuse full-length fluorescent proteins (FPs) in the living nucleus, hampering the application of live cell genomic labeling methods. Here, combining bimolecular fluorescence complementation (BiFC) and transcription activator-like effector (TALE) technologies, we developed a novel method for labeling genomic loci (BiFC-TALE), which largely reduces the background fluorescence level. Using BiFC-TALE, we demonstrated a significantly improved SBR by imaging telomeres and centromeres in living cells in comparison with the methods using full-length FP.