Citation: REN Lin-Jiao, XUE Meng-Xiao, CHEN Qing-Hua, WEI Ming-Hang, ZHANG Pei, QIN Zi-Rui, YAN Yan-Xia, JIANG Li-Ying. Label-free Fluorescent Sensor for Detection of Single-stranded Nucleic Acids Based on G-rich Sequence[J]. Chinese Journal of Analytical Chemistry, ;2023, 51(6): 962-971. doi: 10.19756/j.issn.0253-3820.221546 shu

Label-free Fluorescent Sensor for Detection of Single-stranded Nucleic Acids Based on G-rich Sequence

1.
School of Electrical and Information Engineering, Zhengzhou University of Light Industry, Zhengzhou 450002, China;
2.
College of Computer and Communication Engineering, Zhengzhou University of Light Industry, Zhengzhou 450002, China;
3.
Academy for Quantum Science and Technology, Zhengzhou University of Light Industry, Zhengzhou 450002, China

Corresponding author: CHEN Qing-Hua, JIANG Li-Ying,
Received Date: 3 November 2022
Revised Date: 3 April 2023

Fund Project: Supported by the National Natural Science Foundation of China (Nos. 62073299, 12004348, 61801436), the Project of Central Plains Science and Technology Innovation Leading Talents (No. 224200510026) and the Scientific and Technological Projects in Henan Province, China (Nos. 212102210196, 212102210002, 12102210278, 212102210008).

G-rich sequences with specific conformations (e.g. G-quadruplex and G-triplex) can interact with fluorescent dyes to enhance their fluorescence signal intensity, and are widely used for label-free fluorescent biosensing. In this study, two label-free fluorescent sensors based on G-rich sequences were constructed for detection of the gene sequence of β-amyloid protein (Aβ), a marker of Alzheimer's syndrome, using thiosemicarbazone T (ThT) as the fluorescent dye. The experimental results showed that the G-rich sequences were present as G-triplexes when the length of the hairpin stem was 4 base pairs, and the output signal of the G-triplex sensor decreased with increasing concentration of Aβ gene, with a linear detection range of 1-100 nmol/L and a detection limit of 0.3 nmol/L (S/N = 3). When the length of the hairpin stem was 8 base pairs and the base AATT was added at the 5' end, the G-rich signal decreased with the concentration of Aβ gene. The G-rich sequences were mostly present as G-quadruplexes after binding to Aβ gene, and the output signal of the G-quadruplex sensor was enhanced with increasing concentration of Aβ gene, with a linear detection range of 0.1-100 nmol/L and a detection limit of 0.04 nmol/L (S/N = 3). The two sensors were prepared in a similar process but with different detection principles, providing a basis for further research and application of G-rich sequences. It also provided a new idea for the label-free fluorescence detection of single-stranded nucleic acids.
- G-rich sequence,
- Label-free fluorescence sensor,
- Single-stranded DNA,
- G-triplex,
- G-quadruplex
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沈阳化工大学材料科学与工程学院沈阳 110142