HIV Protocols (4^th Ed., 4th ed. 2024) Methods in Molecular Biology Series, Vol. 2807

Langue : Anglais

Coordonnateurs : Prasad Vinayaka, Kalpana Ganjam

Résumé
Sommaire
Commentaire

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.

Preface…
Table of Contents…
Contributing Authors…

Part I Visualization of HIV-1 RNA and Proteins

1. Infectious Virus Tracking by Fluorescent Live Cell Imaging in Primary Cells
Stephanie Gambut, Thomas J. Hope, and João I. Mamede

2. Imaging HIV-1 Nuclear Import, Uncoating, and Proviral Transcription
Ryan C. Burdick, Alice Duchon, Wei-Shau Hu, and Vinay K. Pathak

3. RNA-FISH for HIV Transcription/Localization Analysis
Pavitra Ramdas and Ajit Chande

4. 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
Rajiv Pathak, Carolina Eliscovich, Robert H. Singer, and Ganjam V. Kalpana

5. Visualizing HIV-1 Assembly at the T Cell Plasma Membrane using Single Molecule Localization: TIRF Microscopy
Rayane Dibsy, Kaushik Inamdar, Cyril Favard, and Delphine Muriaux

6. Single-Virion Analysis: A Method to Visualize HIV-1 Particle Content Using Fluorescence Microscopy
Alice Duchon, Ryan C. Burdick, Vinay K. Pathak, and Wei-Shau Hu

7. Correlative Imaging to Detect Rare HIV Reservoirs and Associated Damage in Tissues
Silvana Valdebenito, David Ajasin, Brendan Prideaux, and Eliseo A. Eugenin

Part II Biomolecular Condensates

8. Characterization of Nuclear HIV-Induced Membraneless Organelles through Fluorescence Microscopy
Viviana Scoca and Francesca Di Nunzio

9. Detection of CPSF6 in Biomolecular Condensates as a Reporter of Nuclear Import
Charlotte Luchsinger and Felipe Diaz-Griffero

Part III HIV-1 Nuclear Import and Chromatin Association

10. Monitoring HIV-1 Nuclear Import Kinetics Using a Chemically Induced Nuclear Pore Blockade Assay
Adarsh Dharan and Edward M. Campbell

11. Biochemical Detection of Capsid in the Nucleus During HIV-1 Infection
Felipe Diaz-Griffero

12. Chromatin Immunoprecipitation of Retroviral Genomes with Antibodies Recognizing Modified Histones and Specific Viral Proteins
Franziska K. Geis and Stephen P. Goff

Part IV HIV-1 RNA - Functions and Modification

13. Measurement of HIV Rev-Rev Response Element Functional Activity
Patrick E. H. Jackson, Godfrey Dzhivhuho, Jing Huang, Marie-Louise Hammarskjold, and David Rekosh

14. Method for the Enrichment of N6-Methyladenosine-Modified Cellular and HIV-1 RNA
Tarun Mishra, Stacia Phillips, and Li Wu

15. Direct Analysis of HIV mRNA m6A Methylation by Nanopore Sequencing
Ethan Honeycutt, Fredrick Kizito, Jonathan Karn, and Thomas Sweet

16. Identifying Individual Pseudouridine (Ψ) Sites Across Transcripts from HIV-1 Infected Cells
Hannah L. Coutts and David G. Courtney

Part V Host-Virus Interactions

17. Proximity Ligation Assay to Detect the Proximity between Host Proteins and Viral Proteins of HIV-1
Sutanuka Chakraborty, Shobith Suresh, Hrimkar Buch, Arun Panchapakesan, and Udaykumar Ranga

Part VI Studies on HIV Neuropathogenesis

18. Modeling HIV-1 Infection in CNS via Infected Monocytes using Immunocompetent Brain Organoids
Roberta S. dos Reis, Sathish Selvam, Marc C.E. Wagner, and Velpandi Ayyavoo

19. An In Vitro Model of Blood Brain Barrier for Studies on HIV Neuroinflammation and CNS Antibody Penetration
Yuekun Lang, Dina Mofed, Vasudev R. Rao, Samantha J. Welninski, Anjelica F. Reyes, Jeffrey R. Schneider, and Vinayaka R. Prasad

Part VII HIV-1 Preclinical Studies

20. Generation of Anti-HIV CAR-T Cells for Preclinical Research
Hang Su, Kim Anthony-Gonda, Rimas J. Orentas, Boro Dropulić, and Harris Goldstein

21. Highly Sensitive Analysis of Cervical Mucosal HIV-1 Infection using Reporter Viruses Expressing Secreted Nanoluciferase
Dana F. Indihar, Jennifer J. Jones, Christina Ochsenbauer, and John C. Kappes

22. Multiparameter Flow Cytometry Monitoring of T Cell Responses
Holden T. Maecker

23. An Ultrasensitive p24 Assay to Measure HIV-1 in Diverse Biological Matrixes
Callie Levinger, J. Natalie Howard, and Alberto Bosque

Subject Index List…