AbstractPromising advances in molecular medicine have promoted the urgent requirement for reliable and sensitive diagnostic tools. Electronic biosensing devices based on field‐effect transistors (FETs) exhibit a wide range of benefits, including rapid and label‐free detection, high sensitivity, easy operation, and capability of integration, possessing significant potential for application in disease screening and health monitoring. In this perspective, the tremendous efforts and achievements in the development of high‐performance FET biosensors in the past decade are summarized, with emphasis on the interface engineering of FET‐based electrical platforms for biomolecule identification. First, an overview of engineering strategies for interface modulation and recognition element design is discussed in detail. For a further step, the applications of FET‐based electrical devices for in vitro detection and real‐time monitoring in biological systems are comprehensively reviewed. Finally, the key opportunities and challenges of FET‐based electronic devices in biosensing are discussed. It is anticipated that a comprehensive understanding of interface engineering strategies in FET biosensors will inspire additional techniques for developing highly sensitive, specific, and stable FET biosensors as well as emerging designs for next‐generation biosensing electronics.