期刊
MOLECULAR THERAPY-NUCLEIC ACIDS
卷 28, 期 -, 页码 450-461出版社
CELL PRESS
DOI: 10.1016/j.omtn.2022.03.022
关键词
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资金
- Swedish Research Council [2018-03075, 2021-02793]
- Swedish Alzheimer Foundation
- Swedish Brain Foundation
- Ahlen Foundation
- Gamla Tjanarinnor Foundation
- Gun and Bertil Stohne's Foundation
- Maximizing Investigators Research Award (MIRA) from the National Institutes of Health [R35 GM118158]
- Desmond and Ann Heathwood MGH Research Scholar Award
- Robert B. Colvin, MD Endowed Chair in Pathology
- Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) [417577129]
- NIH [AG044486, AG015379]
- Swedish Research Council [2018-03075, 2021-02793] Funding Source: Swedish Research Council
The study demonstrates the potential effectiveness of using the CRISPR-Cas9 system to selectively target the PSEN1(M146L) allele and counteract the phenotype associated with early-onset Alzheimer's disease.
Presenilin 1 (PS1) is a central component of gamma-secretase, an enzymatic complex involved in the generation of the amyloid-beta (A beta) peptide that deposits as plaques in the Alzheimer's disease (AD) brain. The M146L mutation in the PS1 gene (PSEN1) leads to an autosomal dominant form of early-onset AD by promoting a relative increase in the generation of the more aggregation-prone A beta 42. This change is evident not only in the brain but also in peripheral cells of mutation carriers. In this study we used the CRISPR-Cas9 system from Streptococcus pyogenes to selectively disrupt the PSEN1(M146L) allele in human fibroblasts. A disruption of more than 50% of mutant alleles was observed in all CRISPR-Cas9-treated samples, resulting in reduced extracellular A beta 42/40 ratios. Fluorescence resonance energy transfer-based conformation and western blot analyses indicated that CRISPR-Cas9 treatment also affects the overall PS1 conformation and reduces PS1 levels. Moreover, our guide RNA did not lead to any detectable editing at the highestranking candidate off-target sites identified by ONE-seq and CIRCLE-seq. Overall, our data support the effectiveness of CRISPR-Cas9 in selectively targeting the PSEN1(M146L) allele and counteracting the AD-associated phenotype. We believe that this system could be developed into a therapeutic strategy for patients with this and other dominant mutations leading to early-onset AD.
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