Biosensors can sensitively and selectively detect a wide range of compounds and macromolecules strongly relevant to human health diagnosis and environment monitoring. Laser induced graphene (LIG) fabricated from polyimide has recently received intense interest for biosensor application due to its unique properties, such as three-dimensional macroporous structure, good conductivity and superior facile laser fabrication process. This laser direct writing technology demonstrates a great potential for developing graphene-based electronics for its chemical-free and direct patterning of graphene, as well as suitability for roll-to-roll production. In this review, we summarize the recent development of the fabrication of LIG and its modification for meeting the needs of biosensor development. The LIG has been directly employed as electrode, modified with enzyme, aptamer or other catalyst for biosensing. The review also highlights integrated LIG biosensors that can simultaneously measure multiple objectives.