Novel Applications and Development of Genome Editing Platforms

Abstract

The discovery of DNA as the primary modality of information storage in living systems has proven to be a watershed moment in our understanding of the underlying mechanisms of biology and disease. Humanity’s expanding grasp of the relationship between DNA and the downstream components of the central dogma, RNA and protein, has proven pivotal in determining why diseases occur and moreover, how to treat them. Only recently, however, has the development of genome editing technologies provided the ability to directly install desired changes in DNA in a precise and programmable manner. This unprecedented capability to probe and manipulate biological systems at a fundamental level has, along with providing new avenues of scientific discovery, raised the theoretical opportunity to cure genetic diseases for the first time. Herein I describe efforts to develop and demonstrate the clinical potential of precision genome editing tools. First, I describe the use of adenine base editors in animal models to treat late-infantile Batten disease. Efficient in vivo base editing-mediated correction of the causative nonsense mutation arrested disease progression in a humanized mouse model of the disease, suggesting high therapeutic potential for such a strategy. I next report the development of engineered prime editing guide RNAs as a general improvement for prime editing, a novel genome editing technology that I assisted in the development of. Through rational analysis and mechanistic studies, we identified exonuclease-mediated degradation as a major bottleneck for prime editing efficiency and developed RNA motifs which mitigate this process. Finally, I describe progress toward the use of prime editing to treat cancer by editing cancer cells to display highly immunogenic epitopes that the human immune system is already poised to attack.