Membrane type-1 matrix metalloproteinase (MT1-MMP) is a type I transmembrane proteinase that has been shown to promote the progression of different diseases, including rheumatoid arthritis (RA) and cancer, by enhancing cellular invasion. MT1-MMP promotes cellular invasion by degrading pericellular extracellular matrix (ECM) at the leading edge of migrating cells. MT1-MMP has been shown to localise to various motility-associated structures, including lamellipodia, filopodia, invadopodia, and podosomes. However, the molecular mechanisms of MT1-MMP localisation to these structures are not well- understood. Dr. Itoh’s research group and others have found that MT1-MMP cell surface exposure is achieved by intracellular trafficking of MT1-MMP-containing vesicles along microtubules and that kinesin superfamily motor proteins (KIFs) play a role. We have employed a siRNA library to screen for the responsible KIFs and found that the knockdown (KD) of four KIFs (KIF13A, KIF3A, KIF9, KIF1C) notably affected MT1-MMP-mediated cellular functions at the cell-matrix interface of human fibrosarcoma cells (HT1080). Interestingly, the KD of these KIFs did not affect the overall level of MT1-MMP at the cell surface, but it significantly influenced MT1-MMP localisation at the substrate-attached side of the plasma membrane. Live cell imaging experiments confirmed that KIF13A, KIF3A and KIF9 associate with MT1-MMP-positive vesicles. These experiments also showed that when cells are cultured atop a substratum, KIF13A and KIF3A collaborate to transport MT1-MMP within the central cytoplasmic areas. However, only KIF13A seemed to reach the cell periphery to deliver the proteinase to the tips of motility-associated structures. Bioinformatic analysis revealed that the expression profile of KIF13A and KIF9 is altered in several cancer types and that these two motor proteins might have cancer-specific roles, partially related to their involvement in MT1-MMP intracellular trafficking. Taken together these findings shed light on KIF-dependent MT1-MMP cell surface exposure and suggest KIF-driven MT1-MMP intracellular trafficking mechanisms as potential targets to control the un-wanted cell invasion that sustains the progression of diseases like RA and cancer.