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Wolfe, Scot
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Academic BackgroundScot Wolfe received his B.S. in Chemistry and Biology from Caltech in 1990, and his Ph.D. from Harvard in Chemistry in 1996. From 1996-2001 he was a post-doctoral fellow at Massachusetts Institute of Technology where his work was supported in part by the Leukemia and Lymphoma Society. In 2001, Dr. Wolfe joined the faculty of UMMS. Engineering precise gene editing systems for application in gene therapy and the analysis of gene regulatory networks My research program is focused on four inter-related areas (see our lab website for more details https://www.umassmed.edu/wolfe-lab/): Creating more precise CRISPR/Cas9 nucleases for gene therapy - We are utilzing our expertise in protein engineering and protein-DNA recognition to create hybrid CRISPR/Cas9 nucleases to achieve single-site cleavage precision within the human genome. Our new gene editing platforms ( Cas9-ZFPs and Cas9-Cas9 fusions) provide dramatic improvements in specificity that make them promising tools to serve as the backbone for new cell-based therapeutics or gene therapy reagents. Utilizing CRISPR/Cas9 nucleases for therapeutic applications - We are utilizing CRISPR-Cas systems to improve the efficiency of genome editing in a variety of different cell types. We are particularly interested in editing in CD34+ hematopoietic stem cells with the goal of therapeutically modifying these cells ex vivo to correct a genetic disorder. These cells would then be returned to the patient through autologous stem cell transplant. We have projects focused on Sickle cell disease, beta-thalassemia and Chronic granulotomous disease. We are also working in collaboration with Charles Emerson (UMMS Wellstone Center) on Limb Girdle Muscular Dystrophy and with Christian Mueller (UMMS-RTI) on Hermansky Pudlak Syndrome. Improving delivery systems for CRISPR/Cas9 nucleases for therapeutic applications - We are collaborating the the Khvorova, Sontheimer and Watts laboratories (UMMS-RTI) to improve delivery systems for CRISPR-Cas nucleases in the context of the NIH Somatic Cell Genome Editing program. Utilizing CRISPR/Cas9 to inactivate latent HIV provirus in reservoir cells - We are collaborating with Jeremy Luban (UMMS – PMM) to lead a team that is investigating the chromatin architecture of latent HIV provirus in reservoir cells. This data will be used to develop precise Cas9 nucleases and corresponding delivery systems to selectively inactivate provirus in reservoir cells to provide a path toward a functional cure for HIV. CRIPSR-Cas9 systems in Zebrafish - We are collaborating with Nathan Lawson (UMMS – MCCB) to develop new and improved CRISPR-Cas tools for genome engineering in zebrafish. We have used ZFNs, TALENs and Cas9 to make targeted knockouts of a number of genes of interest. We are currently working to improve the editing efficiency of CRISPR-based systems for targeted gene knock-outs and knock-ins. Protein-DNA recognition - Our research on protein-DNA recognition is focused primarily on two of the most abundant families of DNA-binding domains in metazoans:Cys2His2 Zinc fingers & Homeodomains
We have performed comprehensive analysis of homeodomain and Cys2His2 zinc finger specificities in D. melanogaster – (fruit fly) in collaboration with Michael Brodsky (UMMS-MCCB) & Gary Stormo (Wash. U). We have coupled this data with specificity data from artificial zinc fingers and homeodomains that we have selected using our bacterial one-hybrid system. Using this information we can build simple qualitative models of recognition that allow the design of homeodomains with novel DNA-binding specificity. This dataset can also be used to broadly predict the specificity of homeodomains ( http://stormo.wustl.edu/flyhd/) and zinc fingers ( http://stormo.wustl.edu/ZFModels/). We continue to explore aspects of DNA recognition by these DNA-binding domain families to both more broadly and accurately predicting the specificity of naturally-occurring family members in all species and for rationally engineering the specificity of these DNA-binding domains for therapeutic development . B1H selection systems - We have developed a bacterial one-hybrid system for rapidly characterizing the DNA-binding specificities of sequence-specific transcription factors, both naturally-occurring and engineered. Using this technology have characterized >50% all of the sequence-specific transcription factors in the D. melanogaster genome ( FlyFactorSurvey) in collaboration with the laboratory of Michael Brodsky (UMMS – MCCB). This dataset is being used to unravel transcription factor regulatory networks within the fly. This selection system can also be used to select Cys2His2 Zinc finger proteins and homeodomains with novel DNA-binding specificity for therapeutic applications.
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Wolfe, Scot