Application of TAL effectors to control gene expression in human cells

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TAL effectors (TALs) from plant-pathogenic bacteria are proteins that act as transcription factors in plant cells and employ a unique DNA-binding domain with programmable specificity. As recently shown by others and us, TAL derivatives also function in human cells and may represent excellent tools for the control of human gene expression. The proposed project aims at optimizing TALs as gene switches in human cells. As targets of TALs we chose elements of medical importance, i.e., the promoter of the interferon beta (IFN- β) gene and the provirus of human immunodeficiency virus (HIV). We will focus on optimizing the parameters that govern TAL efficiency and examine positional effects as well as the effects of TALs on DNA modifications and chromatin structure. Moreover, TALs with different transcription activator and nuclease domains will be tested. We will then analyze the effects of TALs on IFN-β and HIV gene expression. Testing various human virus systems, the consequences of artificial IFN-β expression on the cellular antiviral state will be investigated and the interferon stimulated gene (ISG) expression profiles of cells with viral- and TAL-mediated IFN-β expression will be determined. Targeting the HIV provirus, we aim at applying TALs for activation of HIV from latency and for editing of the provirus, respectively. The goal is to develop the TAL technology for routine applications in human cells and, specifically, for the control of viral infections.