Structure-based Engineering of Next Generation Meganucleases for Genome Editing (360G-Wellcome-204376_Z_16_Z)

Nucleotide repeat disorders comprise some of the most devastating rare diseases in the world. Currently there are no approved treatments for many of these diseases while approved drugs are palliative in nature, outlining an unmet need. A potential therapeutic approach involves engineering naturally occurring homing endonucleases to accurately target a unique sequence containing the edge of the repeat region as well as the repeat itself. This project represents a partnership between Precision Biosciences and Wyatt Yue's laboratory at the Structural Genomics Consortium. The working hypothesis is that the DNA break within the direct repeat will be repaired by single strand annealing (SSA), where the repeat on one side of the break pairs with another copy of the repeat on the other side of the break, resulting in a net reduction in the number of repeats. The project aims to generate a nuclease to target the CTG repeat expansion in the DMPK gene which causes the trinucleotide repeat disorder myotonic dystrophy type 1 (DM1). The partnership will combine structural, biochemical and biophysical methods to characterize the binding of nucleases to the appropriate repeat DNA sequences, generating key information for the development of a therapeutic nuclease for DMl.