| The last decade of biotechnology and medical research was defined by our ability to read the genome; the next decade will be defined by our ability to write it. Precision genome engineering has rapidly advanced thanks to breakthroughs in nucleotide-level DNA targeting using the Transcription Activator-Like Effector Nucleases (TALEN) and Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) - CRISPR-associated protein (Cas) molecular tools. This burgeoning field fundamentally changes the way scientists can interrogate the genome for a plethora of applications, including augmenting our capacity to understand and treat genetic diseases. Chapter I reviews the progress that has been made in precision genome engineering, with a focus on the TALEN system. Chapter II documents materials and methods used throughout this thesis. Chapter III describes a new TALEN system and how it can be used to integrate exogenous DNA into a developing embryo, demonstrating for the first time homology directed repair in vivo. Chapter IV investigates the challenges to editing the mitochondrial genome, the only human-associated genome yet to be edited. Chapter V describes a system for editing the mitochondrial genome in vivo, the first precision engineering platform of the mitochondrial genome. Chapter VI makes an observation about the limitations of precision genome engineering, offers insight into the future of mitochondrial genome editing, and draws parallels between computing and the current state of precision genome engineering. |
