Biofunctionalized conjugated polymers (i.e., carrying enzymes, antibodies, and nucleic acids) are of great interest for many biological applications, yet efficient biofunctionalization of conjugated polymers under biocompatible conditions is challenging. We report a facile strategy to make biofunctionalized conjugated polymers through thiol-ene chemistry with embedded latent disulfide functional groups. This is made possible through the design of a cyclic disulfide-containing dioxythiophene, which can be integrated into a series of conjugated polymers via acid-catalyzed chain-growth polymerization. The utility of such a biofunctionalized polymer with glucose oxidase has been examined in organic electrochemical transistors for the selective sensing of glucose. This work provides a venue for the creation of biofunctional organic semiconductors.