AAV DJ/8-Syn-Cre

A large number of inherited or acquired diseases remain promising targets for human gene therapy. One vector that has shown excellent potential in numerous preclinical and clinical evaluations to date is based on the non-pathogenic adeno-associated virus (AAV). AAV has various properties that make it more attractive than its competitors, such as adenoviral or lentiviral vectors, and one unique advantage is the availability of a large number of natural isolates that vary widely in their properties. The functionality of AAV vector particles is primarily determined by the capsid proteins, and the viral Rep proteins and genome packaging elements are largely interchangeable. Paradoxically, the growing library of naturally occurring and synthetically produced AAV capsid sequences (more than 300 to date) currently poses a dilemma for the rational selection of the best serotype for a specific application. To alter tropism, reduce blocking by neutralizing antibodies or improve transduction efficiency, AAV capsids have been chemically or genetically engineered to generate hybrid capsids by combining properties of multiple serotypes, capsid shuffling, directed evolution, rational mutagenesis or carrying peptide insertions that introduce new receptor binding activities. AAV variants with engineered capsids (AAV2-retro, AAV2g9, AAV-DJ, and AAV-DJ/8) have been used to package CRISPR for transduction in the mouse brain. To provide a higher infection rate across a broad range of tissues than any known natural serotype, AAV-DJ underwent DNA family shuffling to create a hybrid capsid from eight different natural serotypes. AAV-DJ/8 was created by introducing two point mutations in the heparin-binding domain of AAV-DJ to increase brain tissue uptake in vivo, resulting in the ability to infect heart and brain tissues similar to AAV8 and AAV9.