Improved prime editors enable pathogenic allele correction and cancer modelling in adult mice [preprint]
Zhao, Yan G.
Sontheimer, Erik J.
Flotte, Terence R.
Wolfe, Scot A.
UMass Chan AffiliationsGraduate School of Biomedical Sciences
Li Weibo Institute for Rare Diseases Research
Program in Molecular Medicine
Department of Microbiology and Physiological Systems
Department of Pediatrics
Horae Gene Therapy Center
RNA Therapeutics Institute
Department of Molecular, Cell and Cancer Biology
Genetics and Genomics
Molecular, Cellular, and Tissue Engineering
Nucleic Acids, Nucleotides, and Nucleosides
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AbstractPrime 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.
bioRxiv 2020.12.15.422970; doi: https://doi.org/10.1101/2020.12.15.422970. Link to preprint on bioRxiv.
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/29657
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Except where otherwise noted, this item's license is described as The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.