Towards better AAV gene therapy: whole genome siRNA and microRNA high throughput screening for the identification of the molecular determinants governing AAV vector transduction, vector production and vector-induced gene correction

The only hope for the treatment of several inherited disorders is based on gene therapy, that is on the possibility of inserting, into the cells bearing a mutated gene, a normal copy of the same, which might supply a missing function. In some circumstances, the normal gene can even become integrated into the host cell DNA and permanently substitute the disease gene. A very effective vehicle for gene delivery is based on AAV, a small virus with a very simple structure, which has already been used in over 70 different clinical trials, showing promising results. The efficacy of AAV, however, depends strictly upon its interactions with the host cell proteins, which therefore, dictate the fate of the vector once internalized, and its capacity to correct a genetic defect. The main purpose of this project is essentially to understand which human genes regulate AAV efficacy, in order to exploit this information for the generation of improved vectors or treatment modalities. This goal will be met using one of the state-of-the-art methodologies of current biomedical research, based on the possibility of individually inactivating each of the approximately 20,000 human genes, using high-throughput technologies, with the purpose of systematically understanding which are relevant for AAV to function.