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Cell Rep. 2017 Nov 14;21(7):1727-1736. doi: 10.1016/j.celrep.2017.10.075.

CRISPR Transcriptional Activation Analysis Unmasks an Occult γ-Secretase Processivity Defect in Familial Alzheimer's Disease Skin Fibroblasts.

Author information

1
Department of Pathology, Cell Biology and Neurology, Taub Institute, Columbia University Medical Center, 650 West 168th St., New York, NY 10032, USA. Electronic address: keiichiinoue@gakushikai.jp.
2
Department of Pathology, Cell Biology and Neurology, Taub Institute, Columbia University Medical Center, 650 West 168th St., New York, NY 10032, USA.
3
Department of Pathology, Cell Biology and Neurology, Taub Institute, Columbia University Medical Center, 650 West 168th St., New York, NY 10032, USA. Electronic address: asa.abeliovich@gmail.com.

Abstract

Mutations in presenilin (PSEN) 1 and 2, which encode components of the γ-secretase (GS) complex, cause familial Alzheimer's disease (FAD). It is hypothesized that altered GS-mediated processing of the amyloid precursor protein (APP) to the Aβ42 fragment, which is accumulated in diseased brain, may be pathogenic. Here, we describe an in vitro model system that enables the facile analysis of neuronal disease mechanisms in non-neuronal patient cells using CRISPR gene activation of endogenous disease-relevant genes. In FAD patient-derived fibroblast cultures, CRISPR activation of APP or BACE unmasked an occult processivity defect in downstream GS-mediated carboxypeptidase cleavage of APP, ultimately leading to higher Aβ42 levels. These data suggest that, selectively in neurons, relatively high levels of BACE1 activity lead to substrate pressure on FAD-mutant GS complexes, promoting CNS Aβ42 accumulation. Our results introduce an additional platform for analysis of neurological disease.

KEYWORDS:

Alzheimer’s disease; BACE1; CRISPR/Cas9; amyloid precursor protein; fibroblast; induced neuronal cells; presenilin; synergistic activation mediator; β-amyloid; γ-secretase processivity

PMID:
29141208
PMCID:
PMC5704930
DOI:
10.1016/j.celrep.2017.10.075
[Indexed for MEDLINE]
Free PMC Article

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