TY - JOUR T1 - Precision Cas9 Genome Editing <em>in vivo</em> with All-in-one, Self-targeting AAV Vectors JF - bioRxiv DO - 10.1101/2020.10.09.333997 SP - 2020.10.09.333997 AU - Raed Ibraheim AU - Phillip W. L. Tai AU - Aamir Mir AU - Nida Javeed AU - Jiaming Wang AU - Tomás Rodríguez AU - Samantha Nelson AU - Eraj Khokhar AU - Esther Mintzer AU - Stacy Maitland AU - Yueying Cao AU - Emmanouela Tsagkaraki AU - Scot A. Wolfe AU - Dan Wang AU - Athma A. Pai AU - Wen Xue AU - Guangping Gao AU - Erik J. Sontheimer Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/10/09/2020.10.09.333997.abstract N2 - Adeno-associated virus (AAV) vectors are important delivery platforms for therapeutic genome editing but are severely constrained by cargo limits, especially for large effectors like Cas9s. Simultaneous delivery of multiple vectors can limit dose and efficacy and increase safety risks. The use of compact effectors has enabled single-AAV delivery of Cas9s with 1-3 guides for edits that use end-joining repair pathways, but many precise edits that correct disease-causing mutations in vivo require homology-directed repair (HDR) templates. Here, we describe single-vector, ∼4.8-kb AAV platforms that express Nme2Cas9 and either two sgRNAs to produce segmental deletions, or a single sgRNA with an HDR template. We also examine the utility of Nme2Cas9 target sites in the vector for self-inactivation. We demonstrate that these platforms can effectively treat two disease models [type I hereditary tyrosinemia (HT-I) and mucopolysaccharidosis type I (MPS-I)] in mice. These results will enable single-vector AAVs to achieve diverse therapeutic genome editing outcomes.Competing Interest StatementThe authors have filed patent applications on technologies related to this work. G.G. is a scientific co-founder of Voyager Therapeutics, Adrenas Therapeutics, and Aspa Therapeutics and holds equity in these companies. G.G. is an inventor on patents with potential royalties licensed to Voyager Therapeutics, Aspa Therapeutics, and other biopharmaceutical companies. E.J.S. is a co-founder and scientific advisor of Intellia Therapeutics. ER -