Graphene fiber is a macroscopic carbonaceous fiber composed of microscopic graphene sheets, and has attracted extensive attention. Graphene building blocks form a highly ordered structure, resulting in fibers with the same properties as graphene, such as superior mechanical and electrical performance, low weight, excellent flexibility, and ease of functionalization. Moreover, graphene fibers are compatible with traditional textile technologies, facilitating the development of wearable electronics, flexible energy devices, and smart textiles. Graphene fibers were first prepared in 2011 by wet spinning of graphene oxide (GO) solution, which was dispersed in water. Various fabrication methods have been developed to assemble graphene sheets into fibers since then and different strategies have been proposed to optimize their structure and performance. Graphene fibers have applications in numerous fields, including conductors, sensors, actuators, smart textiles, and flexible energy devices. This review aims to provide a comprehensive picture of the preparation approaches, properties, and applications of graphene fibers. Firstly, the preparation processes, unique structures, and properties of three typical carbonaceous fibers-arbon fibers, carbon nanotube (CNT) fibers, and graphene fibers-re compared. It can be seen that graphene fibers possess the unique structures, such as the large grain sizes and highly aligned structure, endowing them with the outstanding properties. Then a variety of fabrication techniques have been summarized, including wet spinning, dry spinning, dry-jet wet spinning, space-confined hydrothermal assembly, film conversion approach, and template-assisted chemical vapor deposition (CVD). Wet spinning is a common method to fabricate high-performance graphene fibers and is promising for the large-scale production of graphene fibers. Besides, various strategies for improving the mechanical, electrical, and thermal properties of graphene fibers are introduced in detail, including well-chosen graphene building blocks, optimized fabrication processes, and high-temperature treatments. Although the electrical and thermal transport properties of typical graphene fibers are better than those of carbon fibers, the strength and modulus of graphene fibers are inferior. Therefore, the enhancement of the mechanical properties of graphene fibers by optimizing the composition of precursors, controlling and adjusting the assembly processes, and exploring feasible post-treatment procedures are essential. Meanwhile, the review outlines the applications of graphene fibers in high-performance conductors, functional fabrics, flexible sensors, actuators, fiber-shaped supercapacitors and batteries. Finally, the persisting challenges and the future scope of graphene fibers are discussed. We believe that graphene fibers will become a new structural and functional material that can be applied in numerous fields in the future, aided by the continuous development of materials and techniques.