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

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.