Since the first discovery of conductive phenomena in polyacetylene, organic semiconductors (OSCs) with the conjugated structure are expected to show their broad prospects as the basic components in various advanced electronic devices in the coming post-silicon era. A large variety of functional OSCs can be obtained by low-cost and simple deposition techniques to exhibit remarkable mechanical flexibility. Hence, they have attracted abundant attention in both academia and industry for decades. Clarifying the structure-property relationship, probing the carrier transport behavior, and realizing high-performance optoelectronic devices with novel functions have been the cornerstones for the sustainable development of organic electronics. In recent years, two-dimensional (2D) OSCs with highly ordered molecular packing and disorder-free structures can be remarkably regarded as the interface-like films, which potentially overcome the bottlenecks of traditional bulk materials. Specifically, OSCs provide an excellent platform for basic researches, and also act as the ideal materials for various emerging optoelectronics, which are promising to bring revolutionary breakthroughs to the micro/nano electronics. In this review article, the recent progresses in the deposition techniques, charge transport behaviors, functional applications, and perspectives with 2D OSCs are discussed. This review aims to provide guidance for the furtherance of fundamental studies, combining organic materials with advanced technologies to promote the development of organic electronics.