University of Massachusetts Medical School Faculty Publications

UMMS Affiliation

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

Publication Date

2020-12-16

Document Type

Article Preprint

Disciplines

Cancer Biology | Disease Modeling | Genetics and Genomics | Molecular Biology | Molecular, Cellular, and Tissue Engineering | Nucleic Acids, Nucleotides, and Nucleosides

Abstract

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.

Keywords

Bioengineering, Prime editors, genome editing

Rights and Permissions

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.

DOI of Published Version

10.1101/2020.12.15.422970

Source

bioRxiv 2020.12.15.422970; doi: https://doi.org/10.1101/2020.12.15.422970. Link to preprint on bioRxiv.

Comments

This article is a preprint. Preprints are preliminary reports of work that have not been certified by peer review.

The PDF available for download is Version 2 of this preprint. The complete version history of this preprint is available at bioRxiv.

Journal/Book/Conference Title

bioRxiv

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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