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Telomere Visualization in Tissue Sections using Pyrrole–Imidazole Polyamide Probes

机译:使用吡咯-咪唑聚酰胺探针在组织切片中观察端粒

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摘要

Pyrrole–Imidazole (PI) polyamides bind to specific DNA sequences in the minor groove with high affinity. Specific DNA labeling by PI polyamides does not require DNA denaturation with harsh treatments of heat and formamide and has the advantages of rapid and less disruptive processing. Previously, we developed tandem hairpin PI polyamide probes (TH59 series), which label telomeres in cultured cell lines more efficiently than conventional methods, such as fluorescence in situ hybridization (FISH). Here, we demonstrate that a TH59 derivative, HPTH59-b, along with immunostaining for specifying cell types in the tissues, visualizes telomeres in mouse and human tissue sections. Quantitative measurements of telomere length with single-cell resolution suggested shorter telomeres in the proliferating cell fractions of tumor than in non-tumor tissues. Thus, PI polyamides are a promising alternative for telomere labeling in clinical research, as well as in cell biology.
机译:吡咯-咪唑(PI)聚酰胺以高亲和力结合小沟中的特定DNA序列。通过PI聚酰胺进行的特定DNA标记不需要通过热和甲酰胺的苛刻处理即可使DNA变性,并且具有快速且破坏性较小的优点。以前,我们开发了串联发夹式PI聚酰胺探针(TH59系列),该探针比传统方法(例如荧光原位杂交(FISH))更有效地标记培养细胞系中的端粒。在这里,我们证明了TH59衍生物HPTH59-b,以及用于指定组织中细胞类型的免疫染色,可以观察到小鼠和人类组织切片中的端粒。用单细胞分辨率对端粒长度进行定量测量表明,与非肿瘤组织相比,肿瘤增殖细胞中的端粒更短。因此,PI聚酰胺是临床研究以及细胞生物学中端粒标记的有前途的替代方法。

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