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HIV-1 provirus excised by single CRISPR/Cas9 RNA guide persists in host cell and may be reactivated

Lai, M.; Maori, E.; Quaranta, P.; Matteoli, G.; Maggi, F.; Sgarbanti, M.; Crucitta, S.; Paci, S.; Turriziani, O.; Freer, G.; Heeney, J. L.; Antonelli, G.; Pistello, M.

2020-11-17 microbiology
10.1101/2020.11.16.384180 bioRxiv
Show abstract

Gene editing may be used to cut out the human immunodeficiency virus type-1 (HIV-1) provirus from the host cell genome and eradicate infection. Here, using cells acutely or latently infected by HIV and treated with long terminal repeat-targeting CRISPR/Cas9, we show that the excised HIV provirus persists for a few weeks and, by means of HIV Integrase, rearranges in circular molecules. Circularization and integration restore proviral transcriptional activity that is enhanced in the presence of exogenous Tat and Rev or tumor necrosis factor-, respectively, in acutely or latently infected cells. Although confirming that gene editing is a powerful tool to eradicate HIV infection, this work highlights that, to achieve this goal, the provirus has to be cleaved in several pieces and the infected cells treated with antiviral therapy before and after editing.

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