Summary
The SPLICER toolbox enables efficient and precise exon skipping by CRISPR base editors, improving the treatment of genetic diseases. It uses a near-PAMless SpCas9 variant and simultaneous editing of splice acceptor and donor sites to reduce aberrant splicing.
Highlights
- SPLICER is a novel toolbox for exon skipping using CRISPR base editors.
- It utilizes a near-PAMless SpCas9 variant to target a broader range of exons.
- Simultaneous editing of splice acceptor and donor sites improves exon skipping efficiency.
- SPLICER reduces aberrant splicing events, including cryptic splicing and intron retention.
- The toolbox has therapeutic potential for treating genetic diseases, such as Alzheimer's disease.
- SPLICER achieves efficient exon skipping in vitro and in vivo, with up to 90% skipping efficiency.
- The toolbox has potential applications in gene therapy and basic research.
Key Insights
- The SPLICER toolbox addresses the limitations of traditional CRISPR-Cas9 systems by utilizing a near-PAMless SpCas9 variant, which enables the targeting of a broader range of exons.
- The simultaneous editing of splice acceptor and donor sites is a key innovation of the SPLICER toolbox, allowing for more efficient and precise exon skipping.
- The reduction of aberrant splicing events, including cryptic splicing and intron retention, is a major advantage of the SPLICER toolbox, as these events can lead to unwanted consequences.
- The SPLICER toolbox has significant therapeutic potential for treating genetic diseases, such as Alzheimer's disease, by enabling the efficient and precise skipping of disease-causing exons.
- The toolbox has been demonstrated to achieve efficient exon skipping in both in vitro and in vivo models, with up to 90% skipping efficiency, highlighting its potential for gene therapy applications.
- The SPLICER toolbox also has potential applications in basic research, such as studying the function of specific exons and their role in disease pathology.
- The use of base editors in the SPLICER toolbox provides a more precise and efficient method for exon skipping compared to traditional CRISPR-Cas9 systems, which can introduce unwanted mutations.
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