UCLA

This work is a culmination of the development of various gene therapy technologies for the treatment of genetic disorders. Chapter 1 highlights the generation of a modified S glycoprotein of SARS-Cov-2 to pseudotype lentiviral vectors which efficiently transduced ACE2-expressing cells with high specificity and contain minimal off-target transduction of ACE2 negative cells. Chapter 2 illustrates the generation of a series of lentiviral vectors using regulatory elements from the SH2D1A locus. By iterative process, we developed two candidate vectors that express SAP protein in the native pattern at levels similar to those of the endogenous gene. These vectors may support reconstitution of the immunologic deficiencies of XLP1 in a safe manner. Chapter 3 delineates an ongoing approach to develop a gene editing strategy for site-specific insertion of a CFTR cDNA cassette for the treatment of Cystic Fibrosis. Utilizing various techniques practiced throughout the laboratory, we designed various improvements to gene editing cargo to increase the cutting efficiency and integration of gene editing cargo including the inclusion of electroporation enhancers, double stranded DNA donor chemical modifications and small molecule inhibitors.