Summary
SPLICER is a highly efficient base editing toolbox that enables in vivo therapeutic exon skipping, improving the treatment of genetic diseases.
Highlights
- SPLICER uses near-PAMless Cas9 nickase variants fused to adenosine or cytosine deaminases for simultaneous editing of splice acceptor and splice donor sequences.
- The toolbox improves exon skipping, reduces aberrant splicing, and enables skipping of exons refractory to single splice site editing.
- SPLICER was demonstrated to reduce the formation of Aβ42 peptides in vitro and enable efficient exon skipping in a mouse model of Alzheimer's disease.
- The toolbox has broad applications in medicine and biotechnology.
- SPLICER overcomes limitations of previous technologies, such as reliance on DNA double-strand breaks and transient effects.
- The study demonstrated the therapeutic potential of SPLICER by targeting APP exon 17, which encodes amino acids that are cleaved to form Aβ plaques in Alzheimer's disease.
- SPLICER reduces the formation of Aβ42 peptides in vitro and enables efficient exon skipping in a mouse model of Alzheimer's disease.
Key Insights
- SPLICER is a highly efficient base editing toolbox that enables in vivo therapeutic exon skipping, improving the treatment of genetic diseases by allowing for the simultaneous editing of multiple splicing elements.
- The use of near-PAMless Cas9 nickase variants fused to adenosine or cytosine deaminases in SPLICER increases the number of exons that can be targeted and enhances editing efficiency at different splice acceptor sites.
- SPLICER reduces aberrant splicing events, including cryptic splicing and intron retention, by disrupting both splice acceptor and splice donor sites, leading to improved exon skipping outcomes.
- The toolbox has the potential to treat genetic diseases by skipping exons that are refractory to single splice site editing, as demonstrated by its ability to skip APP exon 17 in a mouse model of Alzheimer's disease.
- SPLICER provides significant advantages over other methods for exon skipping, such as antisense oligonucleotides, which are currently used as gene therapies for treating Duchenne muscular dystrophy and retinitis pigmentosa.
- The study highlights the importance of extensive characterization of base editing profiles to accomplish efficient exon skipping and minimize unwanted cryptic splicing events.
- The development of SPLICER represents a significant advancement in the field of gene editing, offering a precise and efficient tool for treating genetic diseases by modifying splicing regulatory elements.
Mindmap
Citation
Miskalis, A., Shirguppe, S., Winter, J., Elias, G., Swami, D., Nambiar, A., Stilger, M., Woods, W. S., Gosstola, N., Gapinske, M., Zeballos, A., Moore, H., Maslov, S., Gaj, T., & Perez-Pinera, P. (2024). SPLICER: a highly efficient base editing toolbox that enables in vivo therapeutic exon skipping. In Nature Communications (Vol. 15, Issue 1). Springer Science and Business Media LLC. https://doi.org/10.1038/s41467-024-54529-y