Telomere Visualization in Tissue Sections using Pyrrole-Imidazole Polyamide Probes.
Telomere Visualization in Tissue Sections using Pyrrole-Imidazole Polyamide Probes.
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DOI：
10.1038/srep29261
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发表时间：
2016-07-06
影响因子：
4.6
通讯作者：
Maeshima K
中科院分区：
文献类型：
--
作者：
Sasaki A;Ide S;Kawamoto Y;Bando T;Murata Y;Shimura M;Yamada K;Hirata A;Nokihara K;Hirata T;Sugiyama H;Maeshima K
吡咯-咪唑（PI）聚酰胺以高亲和力结合小沟中的特异性DNA序列。通过PI聚酰胺的特异性DNA标记不需要用热和甲酰胺的苛刻处理使DNA变性，并且具有快速且破坏性较小的加工的优点。在此之前，我们开发了串联发夹PI聚酰胺探针（TH 59系列），它比传统方法，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TH 59衍生物，HPTH 59-B，沿着免疫染色用于指定组织中的细胞类型，可视化小鼠和人类组织切片中的端粒。端粒长度的定量测量与单细胞分辨率表明较短的端粒在肿瘤的增殖细胞组分比非肿瘤组织。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物。
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.