Quantitative Live-Cell Imaging of Human Immunodeficiency Virus (HIV-1) Assembly

Advances in fluorescence methodologies make it possible to
investigate biological systems in unprecedented detail. Over the
last few years, quantitative live-cell imaging has increasingly
been used to study the dynamic interactions of viruses with cells
and is expected to become even more indispensable in the future.
Here, we describe different fluorescence labeling strategies that
have been used to label HIV-1 for live cell imaging and the
fluorescence based methods used to visualize individual aspects of
virus-cell interactions. This review presents an overview of
experimental methods and recent experiments that have employed
quantitative microscopy in order to elucidate the dynamics of late
stages in the HIV-1 replication cycle. This includes cytosolic
interactions of the main structural protein, Gag, with itself and
the viral RNA genome, the recruitment of Gag and RNA to the plasma
membrane, virion assembly at the membrane and the recruitment of
cellular proteins involved in HIV-1 release to the nascent budding
site.