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

Nature communications (2019-01-04)
Jichao Sun, Jared Carlson-Stevermer, Utpal Das, Minjie Shen, Marion Delenclos, Amanda M Snead, So Yeon Koo, Lina Wang, Dianhua Qiao, Jonathan Loi, Andrew J Petersen, Michael Stockton, Anita Bhattacharyya, Mathew V Jones, Xinyu Zhao, Pamela J McLean, Andrew A Sproul, Krishanu Saha, Subhojit Roy
RESUMEN

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.

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Sigma-Aldrich
Anticuerpo anti-APP A4, aa 66-81 de APP {NT}, clon 22C11, clone 22C11, Chemicon®, from mouse
Sigma-Aldrich
Anti-Amyloid Precursor Protein, C-Terminal (751-770) Rabbit pAb, liquid, Calbiochem®
Sigma-Aldrich
Anti-Amyloid beta A4 protein Antibody, clone 2E9, clone 2E9, from rat