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Improved split prime editors enable efficient in vivo genome editing

ABSTRACT: Prime editor (PE) is a precise genome-editing tool capable of all possible base conversions, as well as insertions and deletions without DSBs or donor DNA. The efficient delivery of PE in vivo is critical for realizing its full potential in disease modeling and therapeutic correction. Although PE has been divided into two halves and delivered using dual adeno-associated viruses (AAVs), editing efficiency at different gene loci varies among split sites, and efficient split sites within Cas9 nickase are limited. In this study, by screening multiple split sites, we demonstrated a series of efficient split site when delivering PE by dual-AAV. Additionally, we utilized a feature reported by others recently that RNase could be detached from the Cas9n and designed split sites in the first half of Cas9n. To test the editing efficiency in vivo, a novel dual-AAV split-ePE3 was packaged in AAV9 and delivered via tail vein injection in mice, achieving 24.4% precise genome editing 3 weeks post-injection. Our findings establish an alternative split-PE architecture that could achieve robust gene editing efficiency, facilitating the potential utility both in model organisms and as a therapeutic modality.

ORGANISM(S): Mus musculus

PROVIDER: GSE270813 | GEO | 2024/12/01

REPOSITORIES: GEO

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Improved split prime editors enable efficient in vivo genome editing [Amplicon_Seq_in_vitro]

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2024-12-01 | GSE270854 | GEO

Treatment of a metabolic liver disease by in vivo prime editing in mice

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Project description:Prime editor (PE) has been recently developed to induce efficient and precise on-target editing, whereas its guide RNA (gRNA)-independent off-target effects remain unknown. Here, we used whole-genome and whole-transcriptome sequencing to determine gRNA-independent off-target mutations in cells expanded from single colonies, in which PE generated precise editing at on-target sites. We found that PE triggered no observable gRNA-independent off-target mutation genome-wide or transcriptome-wide in transfected human cells, highlighting its high specificity.

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No observable guide-RNA-independent off-target mutation induced by prime editor [RNA-Seq]

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Genotoxic effects of base and prime editing in human hematopoietic stem cells [RNAseq_BaseEd]

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