Department of Molecular, Cell and Cancer Biology; RNA Therapeutics Institute; Horae Gene Therapy Center; Department of Pediatrics; Department of Microbiology and Physiological Systems; Program in Molecular Medicine; Li Weibo Institute for Rare Diseases Research; Graduate School of Biomedical Sciences
Cancer Biology | Disease Modeling | Genetics and Genomics | Molecular Biology | Molecular, Cellular, and Tissue Engineering | Nucleic Acids, Nucleotides, and Nucleosides
Prime editors (PEs) mediate genome modification without utilizing double-stranded DNA breaks or exogenous donor DNA as a template. PEs facilitate nucleotide substitutions or local insertions or deletions within the genome based on the template sequence encoded within the prime editing guide RNA (pegRNA). However, the efficacy of prime editing in adult mice has not been established. Here we report an NLS-optimized SpCas9-based prime editor that improves genome editing efficiency in both fluorescent reporter cells and at endogenous loci in cultured cell lines. Using this genome modification system, we could also seed tumor formation through somatic cell editing in the adult mouse. Finally, we successfully utilize dual adeno-associated virus (AAVs) for the delivery of a split-intein prime editor and demonstrate that this system enables the correction of a pathogenic mutation in the mouse liver. Our findings further establish the broad potential of this new genome editing technology for the directed installation of sequence modifications in vivo, with important implications for disease modeling and correction.
Bioengineering, Prime editors, genome editing
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DOI of Published Version
bioRxiv 2020.12.15.422970; doi: https://doi.org/10.1101/2020.12.15.422970. Link to preprint on bioRxiv.
P, Liang S, Zheng C, Mintzer E, Zhao YG, Ponnienselvan K, Mir A, Sontheimer EJ, Gao G, Flotte TR, Wolfe SA, Xue W. (2020). Improved prime editors enable pathogenic allele correction and cancer modelling in adult mice [preprint]. University of Massachusetts Medical School Faculty Publications. https://doi.org/10.1101/2020.12.15.422970. Retrieved from https://escholarship.umassmed.edu/faculty_pubs/1872
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