Shondra Pruett-Miller Lab

Genome-editing has been the subject of scientific inquiry for decades. Since its initial development, genome-editing technologies are constantly created and evolving to overcome experimental limitations and technical barriers.

Our lab strives to improve the process of genome-editing. We aim to make these methodologies more robust and more reliable so they can be used as a platform for basic and translational research. 

Eukaryotic cells repair double-stranded breaks through either non-homologous end-joining repair or homology-directed repair (HDR). HDR is a more accurate and definable way to repair a double-stranded break and, thus, a more precise way to edit the genome. We are interested and developing strategies to increase the number of HDR repair events. 

Optimizing targeted deletions

Diseases are often characterized by a particular deletion or translocation event. To create useful model systems to study these diseases, we must be able to edit the genome of our model organism to genetically match that of the disease state. Reliable targeted deletion strategies will allow us to precisely replicate disease genetics in the laboratory and, therefore, allows for the creation of a myriad of model systems. 

Translational applications

Our group is highly collaborative which has allowed the methodologies we develop and optimize to be utilized in a translational capacity. 

Oncogenic fusions are a major driver of cancer in both adults and children, making these cancer-specific hybridized genes attractive targets for therapeutic interventions. In collaboration with the Department of Computational Biology, we used in vitro CRISPR-Cas9 editing in acute lymphoblastic leukemia cell lines to validate oncogenic fusions regulated by alternative splicing. Our contribution has helped narrow the list of the genetic fusions that should be considered for therapeutic targeting.

Hemoglobinopathies 

Hemaglobinopathies are genetically characterized by mutations that change the structure or production of hemoglobin. These mutations provide an attractive target for correcting the pathophysiology of the disease at the level of the gene. 

We are modeling hypotheses, developing disease models and collaborating with the Department of Hematology to develop novel therapeutics for hemoglobinopathies such as Hb SC — a form of sickle cell disease characterized by a Glu-to-Lys modification — and hemoglobin E β-thalassemia — characterized by a single nucleotide substitution in the HBB gene that can cause severe hemolytic anemia when co-inherited with other genetic anomalies. Our work has been instrumental in bringing genome editing to the clinic.