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Author:
Date:
2015-11-20
[Abstract] Telomerase maintains telomeric DNA in eukaryotes during early developments, ~90% of cancer cells and some proliferative stem like cells. Telomeric repeats at the end of chromosomes are associated with the shelterin complex. This complex consists of TRF1, TRF2, Rap1, TIN2, TPP1, POT1 which protect DNA from being recognized as DNA double-stranded breaks. Critically short telomeres or impaired shelterin proteins can cause telomere dysfunction, which eventually induces DNA damage responses at the telomeres. DNA damage responses can be identified by antibodies to 53BP1, gammaH2AX, Rad17, ATM, and Mre11. DNA damage foci at uncapped telomeres are referred to as Telomere dysfunction-Induced Foci (TIFs) (de Lange, 2005; Takai et al., 2003).
The TIF assay is based on the ...
[摘要] 端粒酶在早期发育期间在真核生物中维持端粒DNA，〜90％的癌细胞和一些增殖性干细胞。染色体末端的端粒重复与shelterin复合物相关。该复合物由TRF1，TRF2，Rap1，TIN2，TPP1，POT1组成，其保护DNA不被识别为DNA双链断裂。临界短的端粒或受损的遮蔽蛋白可引起端粒功能障碍，其最终在端粒诱导DNA损伤反应。 DNA损伤反应可以通过抗53BP1，gammaH2AX，Rad17，ATM和Mre11的抗体来鉴定。未封端的端粒的DNA损伤灶被称为端粒功能障碍诱导的Foci（TIF）（de Lange，2005; Takai等人，2003）。
TIF测定基于使用针对遮蔽蛋白如TRF2之一的抗体对抗DNA损伤标记物例如γ-H2AX和端粒的抗体的DNA损伤的共定位检测（Takai等人，2003; de Lange，2002; Karlseder等人，1999）。我们在这里描述的方法可以用于正常的人类和癌细胞。
其他常用的方法 - Telomere限制性片段（TRF）分析（Mender和Shay，2015b）和端粒重复扩增方案（TRAP） ...
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