Individuals with knee joint pathologies exhibit quadriceps dysfunction that, during walking, manifests as smaller peak knee extensor moment (pKEM) and reduced knee flexion excursion. These changes persist despite muscle strengthening and may alter stance phase knee joint loading considered relevant to osteoarthritis risk. Novel rehabilitation strategies that more directly augment quadriceps mechanical output during functional movements are needed to reduce this risk. As an important first step, we tested the efficacy of real-time biofeedback during walking to prescribe changes of ±20% and ±40% of normal walking pKEM values in 11 uninjured young adults. We simultaneously recorded knee joint kinematics, ground reaction forces, and, via ultrasound, vastus lateralis (VL) fascicle length change behavior. Participants successfully responded to real-time biofeedback and averaged up to 55% larger and 51% smaller than normal pKEM values with concomitant and potentially favorable changes in knee flexion excursion. While the VL muscle-tendon unit (MTU) lengthened, VL fascicles accommodated weight acceptance during walking largely through isometric, or even slight concentric, rather than eccentric action as is commonly presumed. Targeted pKEM biofeedback may be a useful rehabilitative and/or scientific tool to elicit desirable changes in knee joint biomechanics considered relevant to the development of osteoarthritis.