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Abstract：
Dead cells always accompany with live cells in vivo and in cell culture. It is important to distinguish dead cells from live cells in various biological studies. Currently, the probes for dead cells are mainly nucleic acid-intercalators, most of which have low affinity and potential toxicity to live cells. In this work, we report a novel aptameric probe (Ch4-1) for the first time, which binds cell nuclei with high affinity (apparent Kd = 6.65 ± 3.40 nM). Ch4-1 was generated by Cell-SELEX process, it was identified to target nucleoproteins in cell nuclei. As an oligonucleotide, Ch4-1 cannot penetrate the integrated cell membrane; therefore, it only binds to dead cells rather than live cells. Compared with traditional DNA-targeting nuclear dyes, Ch4-1 possesses a high affinity to the nucleus, no toxicity to live cells, and can be easily labeled with different fluorescent dyes. It was demonstrated to serve as a probe for distinguishing dead cells from live cells in apoptosis assay, as well as for the nuclear staining of tissue sections.
摘要：
在体内和细胞培养中，死细胞总是伴随着活细胞。在各种生物学研究中，区分死细胞和活细胞是很重要的。目前,死细胞的探针主要是核酸嵌入剂，其中大多数对活细胞具有低亲和力和潜在毒性。在这项工作中,我们首次报道了一种新型的适体探针(Ch4-1),以高亲和力结合细胞核(表观Kd=6.65±3.40nM)。Ch4-1是通过Cell-SELEX过程产生的，它被鉴定为靶向细胞核中的核蛋白。作为寡核苷酸，Ch4-1不能穿透整合的细胞膜；因此，它只与死细胞而不是活细胞结合。与传统的DNA靶向核染料相比，Ch4-1对细胞核具有很高的亲和力，对活细胞没有毒性，并且可以很容易地用不同的荧光染料标记。它被证明是在凋亡测定中区分死细胞和活细胞的探针，以及组织切片的核染色。
