Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, Brno CZ-613 00, Czechia.
Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, Brno CZ-613 00, Czechia; Central European Institute of Technology, Brno University of Technology, Technicka 3058/10, CZ-616 00 Brno, Czechia.
Biosens Bioelectron. 2017 Jun 15;92:133-139. doi: 10.1016/j.bios.2017.01.067. Epub 2017 Feb 9.
Here, we aimed our attention at the synthesis of carbon dots (C-dots) with the ability to interact with DNA to suggest an approach for the detection of DNA damage. Primarily, C-dots modified with amine moieties were synthesized using the one-step microwave pyrolysis of citric acid in the presence of diethylenetriamine. The C-dots showed strong photoluminescence with a quantum yield of 4%. In addition, the C-dots (2.8±0.8nm) possessed a good colloidal stability and exhibited a positive surface charge (ζ=36mV) at a neutral pH. An interaction study of the C-dots and the DNA fragment of λ bacteriophage was performed, and the DNA binding resulted in changes to the photoluminescent and absorption properties of the C-dots. A binding of the C-dots to DNA was also observed as a change to DNA electrophoretic mobility and a decreased ability to intercalate ethidium bromide (EtBr). Moreover, the Förster (or fluorescence) resonance energy transfer (FRET) between the C-dots and EtBr was studied, in which the C-dots serve as an excitation energy donor and the EtBr serves as an acceptor. When DNA was damaged using ultraviolet (UV) radiation (λ=254nm) and hydroxyl radicals, the intensity of the emitted photoluminescence at 612nm significantly decreased. The concept was proved on analysis of the genomic DNA from PC-3 cells and DNA isolated from melanoma tissues.
在这里,我们专注于合成能够与 DNA 相互作用的碳点(C-dots),提出一种用于检测 DNA 损伤的方法。首先,通过柠檬酸在二乙烯三胺存在下的一步微波热解合成了带有胺基的 C-dots。C-dots 具有很强的光致发光性能,量子产率为 4%。此外,C-dots(2.8±0.8nm)具有良好的胶体稳定性,并在中性 pH 值下表现出正表面电荷(ζ=36mV)。我们对 C-dots 和 λ噬菌体 DNA 片段进行了相互作用研究,DNA 结合导致 C-dots 的光致发光和吸收性质发生变化。还观察到 C-dots 与 DNA 的结合导致 DNA 电泳迁移率的变化和嵌入溴化乙锭(EtBr)的能力降低。此外,研究了 C-dots 和 EtBr 之间的Förster(或荧光)共振能量转移(FRET),其中 C-dots 作为激发能量供体,EtBr 作为受体。当使用紫外线(UV)辐射(λ=254nm)和羟基自由基对 DNA 进行损伤时,612nm 处发射的光致发光强度显著降低。该概念通过对 PC-3 细胞的基因组 DNA 和黑色素组织中分离的 DNA 的分析得到了证实。