We are a collaborative group of genome editing scientists at the Translational Neuroscience Center of Boston Children’s Hospital and Harvard Medical School department of Neurology. We strive to develop gene-based therapeutics for genetic neurological diseases. Many neurodegenerative diseases have a genetic underpinning. The cellular processes that result from these genetic abnormalities to cause pathology are often poorly understood. By creative use of genome editing tools, we modify sequences implicated in neurodegenerative diseases, including motor neuron diseases and movement disorders, to study them and correct them at their origin.

There is a wide variety of CRISPR-based genome editing tools available that enable various DNA editing capabilities, each with their own benefits and drawbacks. For example, Cas nucleases are most efficient for generating genetic knockouts and base editing enables precise single-nucleotide changes, whereas prime editing is highly versatile and enables many changes, but is typically less efficient. There are over fifteen different Cas proteins, ten base editor deaminases, and countless pegRNA designs to choose from at a given target locus and it is difficult to predict what exact genome editing strategy will work best for a given application, even by an expert user.  Empirically evaluating the genotypic outcomes and resulting phenotypes of every possible strategy is cumbersome and as a result, typically only a few strategies are tested. This means that better and more efficient genome editing strategies are often not explored.

We study CRISPR-based genome editing tools to deeply understand their interactions with their target substrate by developing high-throughput cellular assays of CRISPR-based genome editing outcomes. These large-scale data improve our understanding of how genome editing tools function, and can be applied to train machine learning models that predict genome editing outcomes to streamline the strategy optimization process. With this improved insight into the functioning of genome editing tools, we can generate atypical edits of functional relevance, and improve precision genome editing of desired targets. For example, we previously demonstrated how Cas-nucleases can be used to correct pathogenic duplication sequences to a wild-type genotype without the use of a homology donor template, and that some base editors can be employed to induce non-canonical nucleotide changes such as C-to-T editing by adenine base editors and transversion edits using cytosine base editors. We can then apply the best tools in the context of genetic neurological diseases for the development of novel genome editing-based therapeutics.

Many motor neuron diseases and movement disorders such as spinal muscular atrophy (SMA), amyotrophic lateral sclerosis (ALS) and ataxias are monogenic and can be corrected or treated by inducing compensatory single nucleotide changes in the genome. Using genome editing, we investigate the impact of genetic mutations of neurological diseases in cells and in animals, and design genome editing approaches to treat or prevent the onset of these diseases. By delivery of genome editing agents in animal models of neurological diseases, we aim to develop novel genome editing-based therapeutics that may one day be used to treat patients.