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Fluorescence spectroscopic detection and measurement of single telomere molecules

机译:单个端粒分子的荧光光谱检测和测量

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摘要

Telomeres are the end-caps of chromosomes that serve to protect the integrity of the genome. Below certain critical lengths, the telomeres can no longer fulfill their protective function, and chromosomal instability ensues. Telomeres shorten during normal cell division due to the end replication problem and are implicated in the development of various aging-associated diseases, including cancer. Telomere length has the potential to serve as a useful biomarker in the field of aging and cancer. However, existing methods of telomere measurement are either too laborious, unable to provide absolute measurement of individual telomere lengths, or limited to certain chromosomes or cell types. Here, we describe an easy single-molecule, fluorescence spectroscopic method for measuring the length of telomeres that permits the profiling of absolute telomere lengths in any DNA sample. We have demonstrated the accurate detection of telomeres as short as 100 bp using cloned telomere standards, and have profiled telomere lengths in human cancer cell lines and primary cells. Since this method allows direct comparison between samples, it could greatly improve the clinical utility of telomere biomarkers.
机译:端粒是染色体的末端,可保护基因组的完整性。在一定的临界长度以下,端粒不能再实现其保护功能,从而导致染色体不稳定。由于末端复制问题,端粒在正常细胞分裂过程中会缩短,并与包括癌症在内的各种衰老相关疾病的发展有关。端粒长度可能在衰老和癌症领域用作有用的生物标记。然而,现有的端粒测量方法要么太费力,不能提供单个端粒长度的绝对测量,要么限于某些染色体或细胞类型。在这里,我们描述了一种简单的单分子荧光光谱方法,用于测量端粒的长度,该方法允许在任何DNA样品中分析绝对端粒的长度。我们已经证明了使用克隆的端粒标准品可以精确检测出短至100 bp的端粒,并在人类癌细胞系和原代细胞中显示了端粒长度。由于这种方法可以直接比较样品,因此可以大大提高端粒生物标记物的临床实用性。

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