Adenine Base Editing in vivo with a Single Adeno-Associated Virus Vector [preprint]
Gao, Xin D.
Wolfe, Scot A.
Sontheimer, Erik J.
UMass Chan AffiliationsGraduate School of Biomedical Sciences
Li 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
Genetics and Genomics
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AbstractBase 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.
bioRxiv 2021.12.13.472434; doi: https://doi.org/10.1101/2021.12.13.472434. Link to preprint on bioRxiv.
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/30750
This article is a preprint. Preprints are preliminary reports of work that have not been certified by peer review.
Related ResourcesNow published in GEN Biotechnology doi: 10.1089/genbio.2022.0015
RightsThe 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.
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.