Single-Cell Imaging of HIV-1 Provirus (SCIP)
Single-Cell Imaging of HIV-1 Provirus (SCIP)
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DOI：
10.1073/pnas.1216254110
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发表时间：
2013-04-02
影响因子：
11.1
通讯作者：
Cereseto, Anna
中科院分区：
文献类型：
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作者：
Di Primio, Cristina;Quercioli, Valentina;Cereseto, Anna
荧光显微镜的最新进展为研究和分析细胞环境中的病毒复制步骤提供了工具。在HIV领域，目前的可视化系统成功地实现了病毒包膜和蛋白质的荧光标记，但不能实现基因组的荧光标记。在这里，我们开发了一个系统，能够可视化的HIV-1的前病毒DNA通过免疫荧光检测修复灶的DNA双链断裂特异性诱导的病毒基因组中的异源表达的I-SceI核酸内切酶。用于HIV-1前病毒单细胞成像的系统，名为SCIP，提供了单独跟踪感染细胞核内整合病毒DNA的可能性。特别是，SCIP使我们能够进行整合的病毒DNA的拓扑分析，揭示HIV-1优先整合在位于细胞核周边的染色质中。
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.