Related Experiment Video
Updated: Mar 14, 2026
Author Spotlight: Optimization of Performance Parameters of the TAGGG Telomere Length Assay
Published on: April 21, 2023
High-throughput identification of telomere-binding ligands based on the fluorescence regulation of DNA-copper
Luzhu Yang1, Yanjun Wang1, Baoxin Li1
1Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an 710062, China.
Abstract:
Formation of the G-quadruplex in the human telomeric DNA is an effective way to inhibit telomerase activity. Therefore, screening ligands of G-quadruplex has potential applications in the treatment of cancer by inhibit telomerase activity. Although several techniques have been explored for screening of telomeric G-quadruplexes ligands, high-throughput screening method for fast screening telomere-binding ligands from the large compound library is still urgently needed. Herein, a label-free fluorescence strategy has been proposed for high-throughput screening telomere-binding ligands by using DNA-copper nanoparticles (DNA-CuNPs) as a signal probe. In the absence of ligands, human telomeric DNA (GDNA) hybridized with its complementary DNA (cDNA) to form double stranded DNA (dsDNA) which can act as an efficient template for the formation of DNA-CuNPs, leading to the high fluorescence of DNA-CuNPs. In the presence of ligands, GDNA folded into G-quadruplex. Single-strdanded cDNA does not support the formation of DNA-CuNP, resulting in low fluorescence of DNA-CuNPs. Therefore, telomere-binding ligands can be high-throughput screened by monitoring the change in the fluorescence of DNA-CuNPs. Thirteen traditional chinese medicines were screened. Circular dichroism (CD) measurements demonstrated that the selected ligands could induce single-stranded telomeric DNA to form G-quadruplex. The telomere repeat amplification protocol (TRAP) assay demonstrated that the selected ligands can effectively inhibit telomerase activity. Therefore, it offers a cost-effective, label-free and reliable high-throughput way to identify G-quadruplex ligands, which holds great potential in discovering telomerase-targeted anticancer drugs.
Insights
A new label-free fluorescence strategy using DNA-copper nanoparticles enables high-throughput screening of telomeric G-quadruplex ligands. This method efficiently identifies compounds that inhibit telomerase activity, offering potential for new cancer drug discovery.
Area of Science:
- Biochemistry
- Molecular Biology
- Nanotechnology
Background:
- G-quadruplex formation in human telomeric DNA inhibits telomerase, a target for cancer therapy.
- Screening for telomeric G-quadruplex ligands is crucial for developing anticancer drugs.
- A high-throughput method for screening telomere-binding ligands is urgently needed.
Purpose of the Study:
- To develop a label-free, high-throughput fluorescence strategy for screening telomere-binding ligands.
- To utilize DNA-copper nanoparticles (DNA-CuNPs) as a signal probe for ligand screening.
- To identify potential anticancer agents targeting telomerase.
Main Methods:
- A label-free fluorescence strategy employing DNA-CuNPs as a signal probe.
- Monitoring fluorescence changes based on G-quadruplex formation induced by ligands.
- Validation using Circular Dichroism (CD) and Telomere Repeat Amplification Protocol (TRAP) assays.
Main Results:
- The DNA-CuNP strategy effectively distinguishes between G-quadruplex binders and non-binders via fluorescence.
- Thirteen traditional Chinese medicines were screened, with selected ligands confirmed to form G-quadruplex structures.
- Selected ligands demonstrated effective inhibition of telomerase activity in TRAP assays.
Conclusions:
- The developed method provides a cost-effective, reliable, and high-throughput approach for identifying G-quadruplex ligands.
- This strategy holds significant potential for discovering novel telomerase-targeted anticancer drugs.
- The findings pave the way for efficient screening of large compound libraries.
