Optimization of base editors for the functional correction of SMN2 as a treatment for spinal muscular atrophy
Brief intro:
- Author: Christiano R. R. Alves, Leillani L. Ha, Rebecca Yaworski, Emma R. Sutton, Cicera R. Lazzarotto, Kathleen A. Christie, Aoife Reilly, Ariane Beauvais, Roman M. Doll, Demitri de la Cruz, Casey A. Maguire, Kathryn J. Swoboda, Shengdar Q. Tsai, Rashmi Kothary & Benjamin P. Kleinstiver
- Journal: Nature Biomedical Engineering
- Doi: https://www.doi.org/10.1038/s41551-023-01132-z
- Publication Date: 2023 Dec 6
Products/Services used in the paper
Quotation shows PackGene:For genome editing experiments, two AAV9 vectors encoding ABE8e-SpRY split into N- and C-terminal fragments via an Npu intein (as described above and similar to a previous report68) paired with gRNA A8 were packaged by PackGene Biotech.
Research Field:spinal muscular atrophy
AAV Serotype:AAV9
Targeted organ:Spinal muscle
Animal or cell line strain:mice
Abstract
Spinal muscular atrophy (SMA) is caused by mutations in SMN1. SMN2 is a paralogous gene with a C•G-to-T•A transition in exon 7, which causes this exon to be skipped in most SMN2 transcripts, and results in low levels of the protein survival motor neuron (SMN). Here we show, in fibroblasts derived from patients with SMA and in a mouse model of SMA that, irrespective of the mutations in SMN1, adenosine base editors can be optimized to target the SMN2 exon-7 mutation or nearby regulatory elements to restore the normal expression of SMN. After optimizing and testing more than 100 guide RNAs and base editors, and leveraging Cas9 variants with high editing fidelity that are tolerant of different protospacer-adjacent motifs, we achieved the reversion of the exon-7 mutation via an A•T-to-G•C edit in up to 99% of fibroblasts, with concomitant increases in the levels of the SMN2 exon-7 transcript and of SMN. Targeting the SMN2 exon-7 mutation via base editing or other CRISPR-based methods may provide long-lasting outcomes to patients with SMA.
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