With various promising applications demonstrated, nanofluidics has been of broad research interest in the past decade. As nanofluidics matures from a proof of concept towards practical applications, it faces two major barriers: expensive nanofabrication and ultra-low throughput. To date, the only material that enables nanofabrication-free, high-throughput, yet precisely controllable nanofluidic systems is the close-packed nanoparticle array, i.e. nanofluidic crystals. Recently, significant progress in nanofluidics has been made using nanofluidic crystals, including high-current ionic diodes, high-power energy harvesters, efficient biomolecular separation, and facile biosensors. Nanofluidic crystals are seen as a key to applying nanofluidic concepts to real-world applications. In this review, we introduce the key concepts and models in nanofluidic crystals, summarize the fabrication methods, and discuss the various applications of nanofluidic crystals in depth, highlighting their advantages in terms of simple fabrication, low cost, flexibility, and high throughput. Finally, we provide our perspectives on the future of nanofluidic crystals and their potential impacts.