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Nat Commun. 2019 Jan 3;10(1):53. doi: 10.1038/s41467-018-07971-8.

CRISPR/Cas9 editing of APP C-terminus attenuates β-cleavage and promotes α-cleavage.

Author information

1
Department of Pathology and Laboratory Medicine, University of Wisconsin-Madison, 1111 Highland Avenue, Madison, WI, 53705, USA.
2
Department of Biomedical Engineering, University of Wisconsin-Madison, 1550 Engineering Drive, Madison, WI, 53706, USA.
3
Wisconsin Institute for Discovery, University of Wisconsin-Madison, 330 N. Orchard, Madison, WI, 53715, USA.
4
Department of Neuroscience, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA, 92093, USA.
5
Waisman Center, University of Wisconsin-Madison, 1500 Highland Ave, Madison, WI, 53705, USA.
6
Department of Neuroscience, Mayo Clinic Jacksonville, 4500 San Pablo Rd, Jacksonville, FL, 32224, USA.
7
Taub Institute for Research on Alzheimer's and the Aging Brain, Columbia University Medical Center, 630W 168th St, New York, NY, 10032, USA.
8
Department of Neuroscience, University of Wisconsin-Madison, 1111 Highland Avenue, Madison, WI, 53705, USA.
9
Department of Pathology and Cell Biology, Columbia University Medical Center, 630W 168th St, New York, NY, 10032, USA.
10
Department of Pathology and Laboratory Medicine, University of Wisconsin-Madison, 1111 Highland Avenue, Madison, WI, 53705, USA. roy27@wisc.edu.
11
Department of Neuroscience, University of Wisconsin-Madison, 1111 Highland Avenue, Madison, WI, 53705, USA. roy27@wisc.edu.

Abstract

CRISPR/Cas9 guided gene-editing is a potential therapeutic tool, however application to neurodegenerative disease models has been limited. Moreover, conventional mutation correction by gene-editing would only be relevant for the small fraction of neurodegenerative cases that are inherited. Here we introduce a CRISPR/Cas9-based strategy in cell and animal models to edit endogenous amyloid precursor protein (APP) at the extreme C-terminus and reciprocally manipulate the amyloid pathway, attenuating APP-β-cleavage and Aβ production, while up-regulating neuroprotective APP-α-cleavage. APP N-terminus and compensatory APP-homologues remain intact, with no apparent effects on neurophysiology in vitro. Robust APP-editing is seen in human iPSC-derived neurons and mouse brains with no detectable off-target effects. Our strategy likely works by limiting APP and BACE-1 approximation, and we also delineate mechanistic events that abrogates APP/BACE-1 convergence in this setting. Our work offers conceptual proof for a selective APP silencing strategy.

PMID:
30604771
PMCID:
PMC6318289
DOI:
10.1038/s41467-018-07971-8
[Indexed for MEDLINE]
Free PMC Article

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