School of Chemistry and Chemical Engineering, Chongqing University of Technology, Chongqing 400054, PR China.
Key Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China.
Spectrochim Acta A Mol Biomol Spectrosc. 2023 Jan 5;284:121760. doi: 10.1016/j.saa.2022.121760. Epub 2022 Aug 13.
Specific and sensitive detection of flap endonuclease 1 (FEN1), an enzyme biomarker involved in DNA replications and several metabolic pathways, is of high values for the diagnosis of various cancers. In this work, a fluorescence strategy based on transcriptional amplification of lighting-up aptamers for label-free, low background and sensitive monitoring of FEN1 is developed. FEN1 cleaves the 5' flap of the DNA complex probe with double flaps to form a notched dsDNA, which is ligated by T4 DNA ligase to yield fully complementary dsDNA. Subsequently, T7 RNA polymerase binds the promoter region to initiate cyclic transcriptional generation of many RNA aptamers that associate with the malachite green dye to yield highly amplified fluorescence for detecting FEN1 with detection limit as low as 0.22 pM in a selective way. In addition, the method can achieve diluted serum monitoring of low concentrations of FEN1, exhibiting its potential for the diagnosis of early-stage cancers.
特异性和灵敏性检测 flap endonuclease 1（FEN1），一种参与 DNA 复制和多种代谢途径的酶生物标志物，对于各种癌症的诊断具有重要价值。在这项工作中，开发了一种基于照明适体转录扩增的荧光策略，用于无标记、低背景和灵敏监测 FEN1。FEN1 切割具有双瓣的 DNA 复合探针的 5' 瓣，形成切口 dsDNA，由 T4 DNA 连接酶连接，生成完全互补的 dsDNA。随后，T7 RNA 聚合酶结合启动子区域，开始循环转录生成许多 RNA 适体，与孔雀石绿染料结合，产生高度放大的荧光，以选择性方式检测 FEN1，检测限低至 0.22 pM。此外，该方法可以实现稀释血清中低浓度 FEN1 的监测，显示其在早期癌症诊断中的潜力。
