Organic mixed ionic-electronic conductors (OMIECs) play a fundamental role in the performance of organic electrochemical transistors (OECTs) and their applications. Although several depletion mode and accumulation mode OMIECs have been utilized for efficient OECT-based glucose sensors, there are still persistent drawbacks such as including biocompatibility, instability, or high detection limits. In this work, a series of indacenodithiophene-based polymeric OMIECs (gIDT, gIDT−T, and gIDT−DTBT) are developed, where the influences of backbone structure on their optical bandgap, energy level, electrochemical propriety, charge transfer and transistor performance, are systematically investigated. By applying KPF 6 electrolyte and vertical device structure, gIDT−DTBT-based vertical OECTs (vOECTs) achieved a maximum output current of −15.63 mA, a maximum transconductance of 39.99 mS, and stable output current (less than ∼2% decay) over 1000 switching cycles. In addition, such vOECTs are employed to detect glucose concentrations ranging from 0.9 to 22.5 μM. A low limit of detection (0.1 μM) and good selectivity are demonstrated. This study indicates that the combination of regulating OMIECs' backbone structure, selecting appropriate electrolytes, and implementing a vertical device structure can help optimize OECT performance and its biosensor applications.