Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, Iran.
Medical Biomaterials Research Center, Tehran University of Medical Sciences, Tehran, Iran.
J Fluoresc. 2017 Jul;27(4):1443-1448. doi: 10.1007/s10895-017-2083-5. Epub 2017 Apr 13.
P53 mutation was detected through the application of a biosensing approach based on the decrease in the fluorescence of oligonucleotide-templated silver nanoclusters (DNA-AgNCs). To this end specific DNA scaffolds of two various nucleotide fragments were used. One of the scaffolds was enriched with two cytosine sequence fragment (C12). This led to DNA-AgNCs with a fluorescence intensity through chemical reduction, while the other scaffold acted as the probe fragment (5- GTAGATGGCCATGGCGCGGACGCGGGTG-3). This latter scaffold selectively bound to the specific p53 site. Thus, resulting AgNCs demonstrated decreased fluorescence upon binding to single-base mismatching targets, and this behavior was found to be linearly proportional to the concentration of mutated p53 from 5 to 350 nM and the approach was found to be able to detect concentrations as low as 1.3 nM.
通过应用基于寡核苷酸模板银纳米簇(DNA-AgNCs)荧光降低的生物传感方法检测到 p53 突变。为此,使用了两种不同核苷酸片段的特定 DNA 支架。其中一个支架富含两个胞嘧啶序列片段(C12)。这导致通过化学还原具有荧光强度的 DNA-AgNCs,而另一个支架则作为探针片段(5-GTAGATGGCCATGGCGCGGACGCGGGTG-3)。后一个支架选择性地与特定的 p53 位点结合。因此,结合单碱基错配靶标后,所得的 AgNCs 的荧光降低,并且发现这种行为与 5 至 350 nM 的突变 p53 浓度呈线性比例,并且该方法能够检测低至 1.3 nM 的浓度。