title: Single-cell imaging of HIV-1 provirus (SCIP).
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-SceI 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.
authors.affiliation: Laboratory of Neurobiology, Scuola Normale Superiore, 56124 Pisa, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Di Primio
authors.forename: Valentina
authors.initials: V
authors.lastname: Quercioli
authors.forename: Awatef
authors.initials: A
authors.lastname: Allouch
authors.forename: Rik
authors.initials: R
authors.lastname: Gijsbers
authors.forename: Frauke
authors.initials: F
authors.lastname: Christ
authors.forename: Zeger
authors.initials: Z
authors.lastname: Debyser
authors.forename: Daniele
authors.initials: D
authors.lastname: Arosio
authors.forename: Anna
authors.initials: A
authors.lastname: Cereseto
mesh_terms: D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D053903:DNA Breaks, Double-Stranded; D017931:DNA Primers / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific; D015497:HIV-1 / Q000648:ultrastructure*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D011533:Proviruses / Q000648:ultrastructure*; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D029701:Saccharomyces cerevisiae Proteins; D059010:Single-Cell Analysis / Q000379:methods
chemical_list: D017931:DNA Primers; D029701:Saccharomyces cerevisiae Proteins; C037725:SCEI protein, S cerevisiae; D015252:Deoxyribonucleases, Type II Site-Specific