Our lab is actively developing CRISPR-based gene-editing nucleases for targeted modification of genomes. We pioneered the development and use of monomeric nuclease platforms based on the GIY-YIG nuclease domain, I-TevI. Currently, we work on development of dual active-site nucleases that introduce precise, defined-length deletions in genomes. These nucleases are based on fusions of the I-TevI nuclease domain to Cas9 and related proteins. Understanding how I-TevI functions as a site-specific monomeric nuclease to change cleavage specificity is an active line of research in the lab.
Publications related to this area of research:
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McMurrough T.A., Brown, C.M., Zhang, Z., Hausner, G., Junop, M.S., Gloor, G.B. and D. R. Edgell (2018) Active site residue identity regulates cleavage preference of LAGLIDADG homing endonucleases. Nucleic Acids Research, 2018 Dec 14;46(22):11990-12007. doi: 10.1093/nar/gky976.
Pereira, C.V., Arguello, T., Bacmam, S.R., Zekonyte, U., Williams, S.L, Edgell, D.R. and C.T. Moraes (2018) MitoTev-TALE: a monomeric DNA editing enzyme to reduce mutant mitochondrial DNA levels in patient-derived cells. EMBO Molecular Medicine, e8084.
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Roy, A.C., Wilson, G.G., and D.R. Edgell (2016) Perpetuating the homing endonuclease life cycle: identification of mutations that modulate and change I-TevI cleavage preference. Nucleic Acids Research, 44 (15): 7350-7359.doi: 10.1093/nar/gkw614.
Wolfs, J.M., Hamilton, T.A., Lant, J.T., Laforet, M., Zhang, J., Salemi, J., Gloor, G.B., Schild-Poulter, C., and D.R. Edgell (2016) Biasing genome-editing events towards precise length deletions with an RNA-guided TevCas9 dual nuclease. Proceedings of the National Academy of Sciences USA, Dec 27;113(52):14988-14993. doi: 10.1073/pnas.1616343114.
Kleinstiver, B.P, Wolfs, J.M., Kolaczyk, T., Roberts, A.K., Hu, S.X., and D.R. Edgell (2012) Monomeric site-specific nucleases for genome editing. Proceedings of the National Academy of Sciences USA 109: 8061-8066.
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