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Figure 1 | Biological Procedures Online

Figure 1

From: A robust dual reporter system to visualize and quantify gene expression mediated by transcription activator-like effectors

Figure 1

Design and Construction of the dual reporters and their functional characterization. (a) Schematic representation of core features of the dual reporter system, depicting the two reporter genes GFP and luciferase separated by T2A (self-cleavage peptide), under the control of a minimal CMV promoter (mCMV). The multiple cloning sites (MCS) were incorporated to facilitate the insertion of TALE binding sequences. (b) Schematic representation of the promoterless dual reporter system serving as a negative control. (c) Schematic representation of the dual reporters under the control of a full CMV promoter serving as a positive control. HEK293 cells transfected with the negative control (promoterless) displayed weak GFP expression (d), while the minimal CMV promoter resulted in low to moderate GFP expression (e); the positive control (full CMV promoter) showed strong GPP expression (f). (g) Luciferase activities of each reporter, displayed in arbitrary units. Error bars indicate SD; n = 3. (h) Flow chart showing how to use the dual reporter to monitor TALE action. Specific TALE reporters can be constructed by inserting the TALE binding sequence (TBS) into the MCS. These customized reporters can then be used to trace TALE action in mammalian cells by co-transfection with the corresponding TALE.

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