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• Reporting synthesis of anisotropic citrate capped gold nanoparticles (ctGNPs) by using curcumin as reducing agent. • The synthesized ctGNPs shows high stability under different physiological condition. • Synthesized ctGNPs using curcumin are non-toxic towards the eukaryotic cells and are more effective towards tumorigenic cell line. • Citrate capped gold nanoparticles (ctGNPs) shows high binding affinity for label-free Human telomeric G-quadruplex DNA due to Van der Waals and H-bonding interactions. We are reporting curcumin-induced synthesis of anisotropic citrate capped Gold nanoparticles (ctGNPs). The techniques such as UV–visible spectroscopy, Raman spectroscopy, FT-IR, X-ray diffraction (XRD), and Transmission Electron Microscopy (TEM) were used to characterize the nanoparticles. The synthetic route shows the formation of an anisotropic gold nanostructure consisting of spherical, triangles, hexagonals, and a low-yield rod with an aspect ratio ranging from 4.2 to 8.5. Curcumin-derived ctGNPs shows stability at different physiological conditions and better biocompatibility. Synthesized nanoparticles are found non-toxic towards eukaryotic cells but more effective against the cancer cell lines HeLa and MCF-7. The interaction of these synthesized nanoparticles with human telomeric G-quadruplex (GQ) DNA was studied using different physiochemical methods. The spectroscopic studies show that synthesized nanoparticles have stronger binding affinity towards telomeric GQ as compared to ds DNA through Van der Waals and H-bonding interactions. Thermodynamic interpretation reveals that the formation of the complex between the telomeric GQ and ctGNPs are enthalpy driven and entropy unfavourable process, resulting in motion freezing and, eventually, AuNP aggregation. Thus, our study shows a new approach to understand the interaction of telomeric G-quadruplexes with gold nanoparticles generated via the green route. [ABSTRACT FROM AUTHOR]