Engineered nucleases for gene therapy

Engineered site-specific endonucleases have emerged today as a promising approach to gene therapy. By targeting specific sequences within the genome, they can stimulate either Homologous Recombination (HR) or Non Homologous End Joining (NHEJ) at a predefined location making possible a wide range of modifications including the correction of mutated alleles, the insertion of a functional coding sequence at a precise location or the generation of functional gene knockouts. We have developed a combinatorial method to entirely redesign the DNA-binding interface of the I-CreI LAGLIDADG meganuclease to create site-specific endonucleases cleaving human genes. Finally, we have compared the activity of engineered meganucleases to that of Transcription Activator-Like Effector Nucleases (TALENs), a novel class of sequence-specific nucleases created by the fusion of transcription activator-like effectors (TALEs) to the catalytic domain of an endonuclease. Data will be presented demonstrating the effective use of both engineered meganucleases and TALENs for genome engineering and will discuss their potential application for therapeutic applications.