Integrase-defective Lentiviral Vectors as a Delivery Platform for Targeted Modification of Adenosine Deaminase Locus.

The disease commonly known as ‘bubble boy disease’ is often caused by a single mutation in the patient’s genome. While there have been gene therapies developed for the disease, a true cure has not been reported. In this study, we aimed towards finding a safe and effective way to reverse the disease-causing mutation. We used a “cut-copy-paste” approach to ‘’correct” the mutations. This was achieved by novel proteins called zinc-finger nucleases, which act as molecular scissors to cut the DNA near the mutation. We use the cell’s own machinery to repair the cut, but using the correct template, to correct the mutation. This method for so called “genome editing” is becoming more and more popular nowadays. But there are still some areas that need to be sorted out. One such area is about delivering these genome-editing reagents to the cells. We looked at integrase-defective lentiviral vectors, which are non-pathogenic viruses that can deliver cargo to cells for a short time. Using the temporarily-delivered nucleases and donor templates, we can achieve genome editing. This study focused on optimizing the use of these vectors, and showed that we can edit the genome at low efficiency. However, there is room for improvement, especially when it comes to using this approach in stem cells. Our studies contribute to the ever-growing body of research on such genome editing strategies.