The early diagnosis of certain fatal diseases is vital for preventing severe consequences and contributes to a more effective treatment. Despite numerous conventional methods to realize this goal, employing nanobiosensors is a novel approach that provides a fast and precise detection. Recently, nanomaterials have been widely applied as biosensors with distinctive features. Graphite phase carbon nitride (g-C₃N₄) is a two-dimensional (2D) carbon-based nanostructure that has received attention in biosensing. Biocompatibility, biodegradability, semiconductivity, high photoluminescence yield, low-cost synthesis, easy production process, antimicrobial activity, and high stability are prominent properties that have rendered g-C₃N₄ a promising candidate to be used in electrochemical, optical, and other kinds of biosensors. This review presents the g-C₃N₄ unique features, synthesis methods, and g-C₃N₄-based nanomaterials. In addition, recent relevant studies on using g-C₃N₄ in biosensors in regard to improving treatment pathways are reviewed.