Titlebook: HIV Protocols ; Vinayaka R. Prasad,Ganjam V. Kalpana Book 2024Latest edition The Editor(s) (if applicable) and The Author(s), under exclus

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Imaging HIV-1 Nuclear Import, Uncoating, and Proviral Transcriptionlution. Very few HIV-1 particles in a viral population are infectious and successfully complete replication (~1/50). Single-particle live-cell imaging enables the study of these rare infectious viral particles, which cannot be accomplished in biochemical assays that measure the average property of t

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RNA-FISH for HIV Transcription/Localization Analysisobservation of messenger RNA (mRNA) molecules associated with particular genes of significance. This technique can also be applied to examine viral transcription and the localization of said transcripts within infected cells. In this context, we provide a comprehensive protocol for the detection, lo

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Single-Cell Single-Molecule RNA-FISH Combined with Immunofluorescence and High-Speed and High-Resolune of the key strategies involves the reactivation of latent HIV-1 from cells using latency-reversing agents. However, currently it is unclear whether any of the latency-reversing agents are able to completely reactivate HIV-1 provirus transcription in all latent cells. An understanding of the react

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Visualizing HIV-1 Assembly at the T-Cell Plasma Membrane Using Single-Molecule Localization Microsco visualization of nanoscaled objects in cell biology. Currently, the use of a recent generation of super-resolution fluorescence microscope coupled with improved fluorescent probes gives the possibility to study the replicative cycle of viruses in living cells, at the single-virus particle or protei

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Correlative Imaging to Detect Rare HIV Reservoirs and Associated Damage in Tissues acquisition, software, spatial resolution, and equipment, including confocal, live-cell, super-resolution, and electron microscopy (scanning, transmission, focused ion beam, and cryo-electron microscopy). However, the recent evolution of different laser-related techniques, such as mass spectrometry

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Characterization of Nuclear HIV-Induced Membraneless Organelles Through Fluorescence Microscopyese steps has a significant impact on HIV overall replication, including integration site selection and viral gene expression. Recently, another important phenomenon has been uncovered as part of HIV interplay with the nuclear environment, specifically involving the cleavage and polyadenylation spec

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Detection of CPSF6 in Biomolecular Condensates as a Reporter of HIV-1 Nuclear Importeus triggers the translocation of CPSF6/CPSF5 from paraspeckles into nuclear speckles, forming puncta-like structures. While this phenomenon is well-documented, the efficiency of CPSF6 translocation to nuclear speckles upon HIV-1 infection varies depending on the type of cell used. In some human cel

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