CRISPR genome editing of HIV-1

Several groups are interested in the possibility of using genome editing to delete or disable the HIV-1 provirus. There are many genome editing technologies, but the most versatile currently seems to be CRISPR. We generated human CD4+ T cell lines expressing CRISPR with a guide RNA targeting different regions of the HIV-1 provirus. We infected the cells with two lab strains of HIV-1 and measured viral replication. As with all mono-therapies, HIV-1 quickly developed strains resistant to CRISPR. We used Next Generation Sequencing to determine the resistance profiles of the viruses. Most HIV-1 resistance to anti-retroviral drugs is due to errors introduced by the viral enzyme reverse transcriptase. We found that this is also true for the CRISPR resistant strains. However, resistance also developed during error-prone DNA repair of the CRISPR induced DNA damage. Thus there are two mechanisms by which HIV-1 can develop resistance to CRISPR genome editing.

Not accepting students for this project.