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Single-Cell Imaging of HIV-1 Provirus (SCIP)

Cristina Di Primio, Valentina Quercioli, Awatef Allouch, Rik Gijsbers, Frauke Christ, Zeger Debyser, Daniele Arosio and Anna Cereseto
Proceedings of the National Academy of Sciences of the United States of America
Vol. 110, No. 14 (April 2, 2013), pp. 5636-5641
Stable URL: http://www.jstor.org/stable/42583035
Page Count: 6
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Single-Cell Imaging of HIV-1 Provirus (SCIP)
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Abstract

Recent advances in fluorescence microscopy provided tools for the investigation and the analysis of the viral replication steps in the cellular context. In the HIV field, the current visualization systems successfully achieve the fluorescent labeling of the viral envelope and proteins, but not the genome. Here, we developed a system able to visualize the proviral DNA of HIV-1 through immunofluorescence detection of repair foci for DNA double-strand breaks specifically induced in the viral genome by the heterologous expression of the I-Scel endonuclease. The system for Single-Cell Imaging of HIV-1 Provirus, named SCIP, provides the possibility to individually track integrated-viral DNA within the nuclei of infected cells. In particular, SCIP allowed us to perform a topological analysis of integrated viral DNA revealing that HIV-1 preferentially integrates in the chromatin localized at the periphery of the nuclei.

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