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Visualization of Genomic Loci in Living Cells with BiFC-TALE

机译:具有BIFC-故事的活细胞中基因组基因座的可视化

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Tracking the dynamics of genomic loci is essential for understanding a variety of cellular processes. However, earlier methods have all suffered from a low signal-to-background ratio (SBR), mainly caused by the background fluorescence from diffuse full-length fluorescent proteins in the nucleus. We have developed a novel method (BiFC-TALE) for labeling and tracking genomic loci in live mammalian cells, combining bimolecular fluorescence complementation (BiFC) and transcription activator-like effector(TALE) technologies. Since only the sequences-targeted BiFC fragments can be pulled together by TALE modules to recombine intact fluorescent proteins, the background fluorescence in the living nucleus can be largely reduced, which significantly improvesSBR. Using telomere and centromere labeling as examples, this unit describes in detail the design and implementation of BiFC-TALE system.
机译:跟踪基因组基因座的动态对于了解各种细胞过程至关重要。 然而,早期的方法均患有低信令对背景比(SBR),主要由核中弥漫性全长荧光蛋白的背景荧光引起。 我们已经开发了一种用于标记和跟踪活哺乳动物细胞中的基因组基因座的新方法(BIFC-轨迹),组合双分子荧光互补(BIFC)和转录活化剂样效应(轨迹)技术。 由于仅通过故事模块将序列靶向的BIFC片段拉到重组完整的荧光蛋白,因此可以在很大程度上降低生命核中的背景荧光,这显着改善了。 使用Telomere和Centromere标签作为示例,本机详细描述了BIFC-TALE系统的设计和实现。

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