Abstract Human Immunodeficiency virus (HIV-1), like all primate lentiviruses possesses the ability to infect non-dividing cells by engaging with components of the nuclear pore complex and mediating the nuclear translocation of the viral ribonucleoprotein complex (RNP) for subsequently integration into the host cell genome. The viral capsid protein (CA) interacts with numerous host factors involving constituents of the nuclear pore complex (NPC) to accomplish the process of nuclear import. Unfortunately, the exact mechanism by which HIV-1 translocates through the nuclear pore complex remains one of the least understood steps of the viral life cycle. In addition, recent evidences supporting the notion of NPCs being more heterogeneous than previously thought further confounds this situation. We have developed an inducible nuclear pore blockade that allows the rate of nuclear import of functional, infectious viral genomes to be monitored in any relevant cells types. Using this technique, we observe that certain CA mutants are insensitive to a Nup62 mediated nuclear pore blockade in cells which potently block infection by wild type CA, demonstrating that HIV-1 can utilize distinct nuclear import pathways during infection. In this application, we will determine the degree to which NPC are heterogeneous and map the specific nuclear pore constituents that makeup the NPCs utilized by HIV-1 during nuclear entry. With our nuclear pore blockade, we now have the capability to block NPCs using different nuclear pore constituents. As such, this application aims to define and visualize the specific nuclear pore constituents that interact and mediate the nuclear import of HIV-1 and how specific NPC usage affects viral integration in target cells. Collectively this application would close critical gaps to our understanding in to the nuclear import of HIV-1.