Molecular Therapy: Nucleic Acids (Jun 2022)

CRISPR-Cas9 treatment partially restores amyloid-β 42/40 in human fibroblasts with the Alzheimer’s disease PSEN1 M146L mutation

  • Evangelos Konstantinidis,
  • Agnieszka Molisak,
  • Florian Perrin,
  • Linn Streubel-Gallasch,
  • Sarah Fayad,
  • Daniel Y. Kim,
  • Karl Petri,
  • Martin J. Aryee,
  • Ximena Aguilar,
  • Bence György,
  • Vilmantas Giedraitis,
  • J. Keith Joung,
  • Vikram Pattanayak,
  • Magnus Essand,
  • Anna Erlandsson,
  • Oksana Berezovska,
  • Martin Ingelsson

Journal volume & issue
Vol. 28
pp. 450 – 461

Abstract

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Presenilin 1 (PS1) is a central component of γ-secretase, an enzymatic complex involved in the generation of the amyloid-β (Aβ) 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 Aβ42. 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 PSEN1M146L 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 Aβ42/40 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 highest-ranking candidate off-target sites identified by ONE-seq and CIRCLE-seq. Overall, our data support the effectiveness of CRISPR-Cas9 in selectively targeting the PSEN1M146L 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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