|dc.description.abstract||Over the past two decades, substantial research has produced a vast array of HIV treatments.
However, the rapid mutation rate of the virus has lead to high drug resistance against the
treatments. To combat the rapid mutations rates, we need to find and design drugs based on
the Host Dependency Factor (HDF)s, the human proteins that are essential for HIV to infect
and replicate. Currently, there is a limited understanding of the how the virus interacts with
the CD4+ T cells at a molecular level. Building on a previous CRISPR/Cas9 whole genome
screen which was conducted to identify these potential HDFs, this study aims to develop a
method to validate the findings.
This was achieved by developing a system with which potential HDFs can be knocked-out
in the cell using CRISPR/Cas9 and subsequently restoring expression via reintroducing the
gene into the genome using a lentiviral vector. This allows us to verify whether any change
in the HIV infection rates is purely due to the single targeted protein. The method developed
in this work was demonstrated on the HIV entry receptor CD4. In this study, we successfully
developed the method on the CD4 surface protein using SUPT1 cells as a T cell model.
The method developed in this study will subsequently be used to validate the identified
HDFs in SUPT1 cells and further in human CD4+ T cells. However, the method can also be
applied to determine any single gene’s affect on a phenotype.||