AAV with CAG2 promoter driven SEAP

This AAV expresses SEAP driven by an ubiquitous CAG2 promoter.

The CAG2 promoter is derived from the commonly used 1.8kb CAG/CBA (also called CAGGS) promoter. To increase the cloning capacity of size-limiting vectors using the CAG promoter, such as AAV or lentiviral vectors, CAG2 was developed by deleting a portion of about 0.6Kb of the chicken beta-actin intron from the original CAG promoter.

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.