Reporter gene constructs

REPORTER GENE CONSTRUCTS WITH VARIABLE AMOUNTS OF 5 -FLANKING DNA... 

Figure 39-11. Location of hormone response elements (HREs) A, B, and C using the reporter gene-transfection approach. A family of reporter genes, constructed as described in Figure 39-10, can be transfected individually into a recipient cell. By analyzing when certain hormone responses are lost in comparison to the 5 deletion, specific hormone-responsive elements can be located.

These type of assays have great potential as screening tools for CYP induction without having to use valuable human hepatocytes. The human hep-atocyte model could be reserved to confirm results of a lead compound after exhaustive screening with such reporter gene construct models. 

Fig. 12.14 Left Structures of mono-substituted alkylpolyamines (AKla-5a), two bri-substituted analogues (AK6b, 7b) were also synthesized and examined for SAR confirmation. Right Inhibition of LPS (100 ng/mL)-induced NF-kB induction in HEK-293 cells stably transfected with Tlr4, MD-2, CD 14, and an NF-icB-secreted alkaline phosphatase reporter gene construct. Polymyxin B was used as the positive control...

Regardless of the expression of the receptor and cofactors, the reporter gene must be introduced to the cells. There are several options for the introduction of the reporter gene construct into the cells including chemical-based transfection (CaClj), lipid-based transfection, and electroporation. 

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