Abstract: The human immunodeficiency virus (HIV) integrates its genome into that of the human host to cause a persistent infection that can be controlled by antiviral drugs, but that cannot be cured. CRISPR-Cas endonucleases can be instructed to cleave and inactivate the integrated proviral DNA genome. We observed many surprise findings along this route. For instance, it turned out that HIV escape is promoted by the cellular DNA repair machinery, which introduces small indels at the target site that trigger subsequent viral escape. When two HIV targets are attacked to realize excision of a large HIV segment, we observed instead that a regular indel is introduced at both targets. To explain this, we reasoned that the two cuts are not present simultaneously because of fast DNA repair. Novel kinetic experiments support this idea. Another surprise finding is that we can score a fair number of very large genome deletions that extend beyond the integrated HIV genome and that are potentially dangerous because of oncogene activation. Most studies will miss such products because of the used PCR-based detection methods. We will also discuss some technological developments, e.g. the development of novel expression cassettes for guide RNAs, Pol-III driven transcription units and novel lentivirus vector designs.

Keywords: HIV, CRISPR-Cas, Cure

Biography: Ben Berkhout studied molecular biology at Leiden University and obtained his PhD in 1986 on translational control by means of RNA structure in bacteriophages. He performed postdoctoral research at the Dana Farber Cancer Institute (Harvard Medical School) and the National Institutes of Health (USA). He became Head of the Laboratory of Experimental Virology and was appointed Professor of Human Retrovirology the University of Amsterdam in 2002. He supervised 55 PhD students and published over 600 manuscripts on HIV-1 replication, virus evolution, virus discovery and new antiviral therapeutic strategies. BB sits on many international science panels (e.g. ERC, RGC Hong Kong, NMRC Singapore), and is editor-in-chief of Virus Research and editor for several journals (e.g. Retrovirology).