GFP Adeno-Associated Virus ( AAV-EARVRPP )

AAV-based recombinant viral vectors have several beneficial properties for gene therapy due to their lack of pathogenicity, high virion stability, and relatively low immunogenicity. Although the virus is primarily extrachromosomal and not suitable for long-term expression in rapidly dividing tissues, it is well suited for "hit and run" applications in these cell types (e.g., drug or radioresistance gene transfer, especially gene correction by homologous recombination) without the potential hazards associated with integration (insertional mutagenesis) and long-term exposure to nonphysiological transgene levels. rAAV2 vectors have been widely used in many clinical and preclinical studies, such as the treatment of coagulation factor disorders, cystic fibrosis, and several types of cancer. Müller and colleagues generated rAAV capsid mutants that provide increased gene transfer efficiency and potentially higher target cell specificity. For this purpose, an AAV random peptide library was used that displays a random seven-amino acid peptide sequence within the VP capsid protein domain (at Arg588), which is typically required for AAV2 binding to one of its natural receptors, heparan sulfate. When selecting the AAV random peptide library on target cells, mutants with good binding properties to the target receptor are able to transduce cells, replicate, and propagate in further rounds of selection. These mutants may exhibit increased transduction efficiency and/or increased specificity for target cells, which must be evaluated in further assays.