Transcriptional Activators of Human Genes with Programmable DNA-Specificity (original) (raw)
Figure 4
Designed TAL proteins induce expression of human genes.
Artificial TAL proteins (HuTALs) containing a VP16-AD were constructed that target promoter regions of human PUMA, IFNα1, or IFNβ1 genes. (A) Target DNA boxes were cloned upstream of the minimal CMV promoter into the luc reporter vector. Luc reporter constructs were co-transfected with constructs driving TAL gene expression into human Hek293T-Rex cells. Measurements were done 1 dpi (error bars indicate SD). Luc activity was plotted in arbitrary light units per µg protein. (B) TAL protein-directed expression of endogenous human genes. Plasmids expressing HuTALs or Hax3 were transfected into human Hek293T-Rex cells. mRNAs levels of target genes were determined by quantitative real-time PCR. mRNA levels in presence of Hax3 (negative control, white bars) were set to 1. (C) The natural IFNβ1 promoter was cloned into the luc reporter vector. The IFNβ1 reporter was co-transfected with constructs directing expression of HuTALIFNβ1 or hax3, respectively. Dark grey bars: HuTAL, white bars: Hax3. (A,C) P≤0.005 for all values; (B) PUMA, P = 0.0051; IFNα1, P = 0.11; IFNβ1, P = 0.00028.