Adenine Base Editing in vivo with a Single Adeno-Associated Virus Vector [preprint]
Authors
Zhang, HanBamidele, Nathan
Liu, Pengpeng
Ojelabi, Ogooluwa
Gao, Xin D.
Rodríguez, Tomás
Cheng, Haoyang
Xie, Jun
Gao, Guangping
Wolfe, Scot A.
Xue, Wen
Sontheimer, Erik J.
UMass Chan Affiliations
Graduate School of Biomedical SciencesLi Weibo Institute for Rare Diseases Research
Department of Microbiology and Physiological Systems
Viral Vector Core
Horae Gene Therapy Center
Program in Molecular Medicine
Department of Molecular, Cell and Cancer Biology
RNA Therapeutics Institute
Document Type
PreprintPublication Date
2022-02-07Keywords
AAVbase editing
genome editing
CRISPR
sgRNA
deaminase
gene therapy
Genetics and Genomics
Molecular Biology
Therapeutics
Viruses
Metadata
Show full item recordAbstract
Base editors (BEs) have opened new avenues for the treatment of genetic diseases. However, advances in delivery approaches are needed to enable disease targeting of a broad range of tissues and cell types. Adeno-associated virus (AAV) vectors remain one of the most promising delivery vehicles for gene therapies. Currently, most BE/guide combinations and their promoters exceed the packaging limit (~5 kb) of AAVs. Dual-AAV delivery strategies often require high viral doses that impose safety concerns. In this study, we engineered an adenine base editor using a compact Cas9 from Neisseria meningitidis (Nme2Cas9). Compared to the well-characterized Streptococcus pyogenes Cas9-containing ABEs, Nme2-ABE possesses a distinct PAM (N4CC) and editing window, exhibits fewer off-target effects, and can efficiently install therapeutically relevant mutations in both human and mouse genomes. Importantly, we show that in vivo delivery of Nme2-ABE and its guide RNA by a single-AAV vector can efficiently edit mouse genomic loci and revert the disease mutation and phenotype in an adult mouse model of tyrosinemia. We anticipate that Nme2-ABE, by virtue of its compact size and broad targeting range, will enable a range of therapeutic applications with improved safety and efficacy due in part to packaging in a single-vector system.Source
bioRxiv 2021.12.13.472434; doi: https://doi.org/10.1101/2021.12.13.472434. Link to preprint on bioRxiv.
DOI
10.1101/2021.12.13.472434Permanent Link to this Item
http://hdl.handle.net/20.500.14038/30750Notes
This article is a preprint. Preprints are preliminary reports of work that have not been certified by peer review.
Related Resources
Now published in GEN Biotechnology doi: 10.1089/genbio.2022.0015Rights
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
10.1101/2021.12.13.472434
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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.