Adenoviral vectors as genome editing tools : repairing defective DMD alleles

Abstract

Adenoviral vectors (AdVs) constitute powerful gene delivery vehicles. However, so far, their potential for genome editing has not been extensively investigated. By tailoring AdVs as carriers of designer nucleases and donor DNA sequences, the research presented in this thesis expands the utility of the AdV platform to genome editing. In particular, in the first part of this thesis, AdVs are exploited for tackling two of the major bottlenecks of genome editing: (i) developing improved methods for delivering the sizable gene-editing tools, such as RNA-guided nuclease complexes, into target cells, and (ii) increasing the specificity and fidelity of the gene-editing procedures. In the second part of the thesis, the insights derived from these studies are further exploited for testing AdVs encoding nucleases as repairing agents of defective DMD alleles in muscle cell populations derived from patients with Duchenne muscular dystrophy (DMD). Finally, the application of AdVs as gene editing tools for repairing endogenous DMD alleles is discussed in the context of other viral vector-based DMD editing strategies. Taken together, the findings reported in this work are expected to aid in the designing and testing of new therapeutic interventions for tackling DMD and are anticipated to be applicable to other genetic disorders.