G-quadruplex is a unique DNA secondary structure formed by guanine-rich nucleic acid sequences. Synonymous terms for G-quadruplex include G4 DNA, G-tetrad, quadruplex DNA, guanine tetrads, and tetraplex DNA. They are characterized by the presence of stacked guanine quartets held together by Hoogsteen hydrogen bonding. Unlike traditional DNA double helix, G-quadruplexes have four-stranded architectures and are formed in regions of the genome that are rich in guanine. G-quadruplexes play essential roles in gene regulation, DNA replication, telomere maintenance, and genome stability. The discovery of G-quadruplexes has revolutionized our understanding of DNA structure and function, and their study holds tremendous therapeutic potential in the treatment of various diseases including cancer.