Improved prime editors enable pathogenic allele correction and cancer modelling in adult mice [preprint]
Authors
Liang, Shun-QingZheng, Chunwei
Mintzer, Esther
Zhao, Yan G.
Ponnienselvan, Karthikeyan
Mir, Aamir
Sontheimer, Erik J.
Gao, Guangping
Flotte, Terence R.
Wolfe, Scot A.
Xue, Wen
UMass Chan Affiliations
Graduate School of Biomedical SciencesLi 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
Document Type
PreprintPublication Date
2020-12-16Keywords
BioengineeringPrime editors
genome editing
Cancer Biology
Disease Modeling
Genetics and Genomics
Molecular Biology
Molecular, Cellular, and Tissue Engineering
Nucleic Acids, Nucleotides, and Nucleosides
Metadata
Show full item recordAbstract
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.Source
bioRxiv 2020.12.15.422970; doi: https://doi.org/10.1101/2020.12.15.422970. Link to preprint on bioRxiv.
DOI
10.1101/2020.12.15.422970Permanent Link to this Item
http://hdl.handle.net/20.500.14038/29657Notes
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 Preprints are preliminary reports of work that have not been certified by peer review.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.
Rights
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.Distribution License
http://creativecommons.org/licenses/by-nc-nd/4.0/ae974a485f413a2113503eed53cd6c53
10.1101/2020.12.15.422970
Scopus Count
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.