AAV with GFAP(0.7) promoter driven SEAP

This AAV expresses SEAP driven by an astrocyte GFAP(0.7) promoter.

This 0.7 Kb novel GFAP promoter was constructed by ligating several key regulatory elements of the 2.2Kb human GFAP promoter. It drives essentially the same expression pattern as the parental 2.2 Kb GFAP promoter but with two-fold greater activity.

The SEAP reporter gene encodes a truncated form of the human placental alkaline phosphatase that lacks the membrane anchoring domain. Therefore, the protein can be efficiently secreted from transfected cells allowing for detection of reporter gene activity without cell lysis. Using a secreted reporter protein has several advantages over traditional reporter assays: 1) Cell lysis is not required for analysis so a single set of cells can be used for both the SEAP assay and another purpose; 2) Gene expression kinetics can be studied by the repeated collection of the culture medium from the same cultures; and 3) By changing the culture medium prior to an experiment, the assay background is reduced to an extremely low level.