CRISPR/Cas9 Ablation of Integrated HIV-1 Accumulates Proviral DNA Circles with Reformed Long Terminal Repeats
Michele Lai, Eyal Maori, Paola Quaranta, Giulia Matteoli, Fabrizio Maggi, Marco Sgarbanti, Stefania Crucitta, Simone Pacini, Ombretta Turriziani, Guido Antonelli, Jonathan L. Heeney, Giulia Freer, Mauro Pistello
Abstract
. Among the different approaches, CRISPR/Cas9 is the most promising tool for gene editing. The present study underlines the remarkable effectiveness of CRISPR/Cas9 in removing the HIV-1 provirus from infected cells and investigates the fate of the excised HIV-1 genome. This study demonstrates that the free provirus may persist in the cell after editing and in appropriate circumstances may reactivate. As an episome, it might be transcriptionally active, especially in the presence of Tat and Rev. The persistence of the HIV-1 episome was strongly decreased by gene editing with multiple targets. Although gene editing has the potential to eradicate HIV-1 infection, this work highlights a potential issue that warrants further investigation.