Full Description
This fourth edition volume expands on the previous editions with discussions on the latest methodologies to study HIV, live cell imaging, HIV cure, new modifications to the viral RNA that impacts HIV biology, and new types of intracellular compartments. The chapters in this book are organized into seven parts and cover topics such as HIV latency reactivation via single molecule RNA detection; T cell responses; new and efficacious anti-HIV CAR T cells; analysis of mucosal HIV infection; analysis of 3D brain organoids to study neuro AIDS; and the transfer of antibodies across the blood brain barrier. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and authoritative, HIV Protocols, Fourth Edition is a valuable resource for all preclinical HIV-1 researchers looking to learn more about this important and advancing field.
Contents
Infectious Virus Tracking by Fluorescent Live Cell Imaging in Primary Cells.- Imaging HIV-1 Nuclear Import, Uncoating, and Proviral Transcription.- RNA-FISH for HIV Transcription/Localization Analysis.- Single-Cell and Single-Molecule RNA-FISH Combined with Immunofluorescence and High-Speed and High-Resolution Scanning Analysis to Visualize Reactivation of Latent HIV-1.- Visualizing HIV-1 Assembly at the T Cell Plasma Membrane using Single Molecule Localization: TIRF Microscopy.- Single-Virion Analysis: A Method to Visualize HIV-1 Particle Content Using Fluorescence Microscopy.- Correlative Imaging to Detect Rare HIV Reservoirs and Associated Damage in Tissues.- Characterization of Nuclear HIV-Induced Membraneless Organelles through Fluorescence Microscopy.- Detection of CPSF6 in Biomolecular Condensates as a Reporter of Nuclear Import.- Monitoring HIV-1 Nuclear Import Kinetics Using a Chemically Induced Nuclear Pore Blockade Assay.- Biochemical Detection of Capsid in the Nucleus During HIV-1 Infection.- Chromatin Immunoprecipitation of Retroviral Genomes with Antibodies Recognizing Modified Histones and Specific Viral Proteins.- Measurement of HIV Rev-Rev Response Element Functional Activity.- Method for the Enrichment of N6-Methyladenosine-Modified Cellular and HIV-1 RNA.- Direct Analysis of HIV mRNA m6A Methylation by Nanopore Sequencing.- Identifying Individual Pseudouridine (Ψ) Sites Across Transcripts from HIV-1 Infected Cells.- Proximity Ligation Assay to Detect the Proximity between Host Proteins and Viral Proteins of HIV-1.- Modeling HIV-1 Infection in CNS via Infected Monocytes using Immunocompetent Brain Organoids.- An In Vitro Model of Blood Brain Barrier for Studies on HIV Neuroinflammation and CNS Antibody Penetration.- Generation of Anti-HIV CAR-T Cells for Preclinical Research.- Highly Sensitive Analysis of Cervical Mucosal HIV-1 Infection using Reporter Viruses Expressing Secreted Nanoluciferase.- Multiparameter Flow Cytometry Monitoring of T Cell Responses.- An Ultrasensitive p24 Assay to Measure HIV-1 in Diverse Biological Matrixes.
