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dc.contributor.advisorMeås, Hany Zakaria
dc.contributor.advisorBösl, Korbinian
dc.contributor.advisorHaug, Markus
dc.contributor.authorBaskaran, Joshua
dc.date.accessioned2021-09-25T16:33:58Z
dc.date.available2021-09-25T16:33:58Z
dc.date.issued2020
dc.identifierno.ntnu:inspera:56877753:34517408
dc.identifier.urihttps://hdl.handle.net/11250/2783279
dc.descriptionFull text not available
dc.description.abstract
dc.description.abstractOver 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.
dc.language
dc.publisherNTNU
dc.titleDevelopment of a CRISPR/Cas9 knockout-complementation system for application in HIV host dependency factor analysis.
dc.typeMaster thesis


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