Ligand coupling to the AAV capsid for cell-specific gene transfer

Abstract

Cell entry of AAV vectors is initiated by contacting the cell surface attachment receptor. This process can be rationally engineered through mutating the contact residues on the AAV capsid and covalently coupling targeting ligands to the capsid surface that exhibit high affinity for a cell surface protein of choice. This way, selective gene delivery to target-receptor positive cell types has been achieved. Two methods for coupling targeting ligands to the AAV capsid can be distinguished. Genetic coupling is achieved through expressing fusion proteins composed of the capsid protein VP2 and the targeting ligand in packaging cells. Biochemical coupling involves split-intein-mediated protein trans-splicing between the mutated AAV capsid and the targeting ligand. While genetic coupling is restricted to designed ankyrin repeat proteins as targeting ligand, biochemical coupling tolerates single-chain antibody fragments as well.