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This research aims to develop a protocol that reverses HIV-1 latency using engineered bacteriophage T4 nanoparticles.

July 20, 2026STAR Protocols0 citationsOpen Access

Protocol for HIV-1 latency reversal using engineered bacteriophage T4 particles

Key Points

This research aims to develop a protocol that reverses HIV-1 latency using engineered bacteriophage T4 nanoparticles.
Prepare CD4-DARPin-T4 nanoparticles using E. coli.
Decorate T4 capsid nanoparticles with CD4-targeting DARPin ligands.
Assess proviral reactivation with fluorescence readouts.
Successfully produced T4 nanoparticles that target CD4+ T cells.
Demonstrated effective proviral reactivation in an HIV-1 latency model.
Provided a clear protocol for producing and utilizing engineered nanoparticles.

Abstract

A major barrier to curing HIV-1 is the latent viral reservoir in resting CD4+ T cells. Here, we present a protocol for preparing CD4-DARPin (designed ankyrin repeat protein)-T4 nanoparticles and their application to HIV-1 latency reversal in a model of HIV-1 latency. We describe steps for producing T4 capsid nanoparticles in E. coli and their decoration with the CD4-targeting DARPin ligand fused to the T4 outer capsid protein Hoc. We then detail procedures for assessing proviral reactivation using fluorescence readouts. For complete details on the use and execution of this protocol, please refer to Batra et al. 1

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