Nuclease resistance of telomere-like oligonucleotides monitored in live cells by fluorescence anisotropy imaging
Nuclease resistance of telomere-like oligonucleotides monitored in live cells by fluorescence anisotropy imaging
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DOI：
10.1021/ac0517601
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发表时间：
2006-03-01
影响因子：
7.4
通讯作者：
Tan, WH
中科院分区：
文献类型：
--
作者：
Cao, ZH;Huang, CC;Tan, WH
端粒具有重要的生物学功能，如保护染色体。在本文中，我们开发了一种荧光各向异性成像系统，用于监测活细胞内DNA的消化。研究了端粒样ssdna在人乳腺癌细胞核中的抗核酸能力。在实验过程中，我们发现这些寡核苷酸明显比常规DNA序列更稳定。我们得出结论，端粒样ssDNA的g -四重结构使其在细胞内环境中比非g -四重结构更稳定。这将有助于我们理解为什么几乎所有真核细胞都采用g -四重体形成的端粒序列来保护染色体的末端。这是第一次在活细胞中观察到这种现象。我们的荧光各向异性成像提供了一种有效的方法，可以实时直接监测活细胞任何区域的DNA消化，为许多重要和相关的细胞内过程提供见解。
Telomeres carry important biological functions such as the protection of chromosomes. In this paper, we have developed a fluorescence anisotropy imaging system for monitoring DNA digestion inside live cells. The nuclease-resistant capability of telomere-like ssDNAs in nuclei of human breast cancer cells is studied. We found that those oligonucleotides were clearly more stable than regular DNA sequences during the time course of the experiments. We conclude that the G-quadruplex structure of the telomere-like ssDNA makes it inherently more stable in intracellular environments than non-G-quadruplex structures. This will help us understand why the G-quadruplex forming telomere sequences were adopted by almost all eukaryotic cells to protect the ends of chromosomes. This is the first time such a phenomenon was observed in live cells. Our fluorescence anisotropy imaging provides an efficient way to directly monitor DNA digestion in any region of live cells in real time, providing insights into many important and related intracellular processes.