Studying viral protein-primed DNA replication to develop new gene editing technologies

There are over 6,000 human genetic diseases, affecting hundreds of millions of people. The vast majority are caused by small mutations in DNA sequences which are potentially correctable, so the development of new gene editing tools will have major benefits for treating human genetic diseases. To tackle this challenge, we will harness the natural protein diversity of viruses to create new gene editing technologies.We will study an unusual class of viruses that use a unique mechanism to replicate themselves, known as protein-primed DNA replication, which creates the rare feature of a protein-DNA covalent bond. These viruses are very understudied, yet they hold great potential for new biotechnology applications. We will study the protein-primed DNA replication machinery of these viruses, and then exploit this mechanism to develop a novel technology for synthetic biology – creating new ways to amplify and deliver DNA into living cells for gene editing.Our research represents a major breakthrough on two accounts:Firstly, to discover and characterise large numbers of protein-primed DNA replication proteins from previously unstudied viruses; to establish a highly efficient, self-replicating system able to synthesise and amplify large amounts of protein-linked DNA product; and to study the molecular details of these viral replication proteins for the first time.Secondly, to develop protein-linked DNA as a new platform technology for gene editing. Our strategy to engineer viral proteins will create protein-linked DNA molecules that are for the first time (i) actively nuclear-targeted and (ii) self-replicating, to address key limitations in the gene editing field and pioneer a new method for highly efficient homology-directed repair.Overall, our proposal combines fundamental biological study and applied biotechnology research to transform our understanding of these viral proteins and engineer them for ground-breaking advances in gene editing and DNA delivery.