TY - JOUR
T1 - CRISPR/Cas9 editing of APP C-terminus attenuates β-cleavage and promotes α-cleavage
AU - Sun, Jichao
AU - Carlson-Stevermer, Jared
AU - Das, Utpal
AU - Shen, Minjie
AU - Delenclos, Marion
AU - Snead, Amanda M.
AU - Koo, So Yeon
AU - Wang, Lina
AU - Qiao, Dianhua
AU - Loi, Jonathan
AU - Petersen, Andrew J.
AU - Stockton, Michael
AU - Bhattacharyya, Anita
AU - Jones, Mathew V.
AU - Zhao, Xinyu
AU - McLean, Pamela J.
AU - Sproul, Andrew A.
AU - Saha, Krishanu
AU - Roy, Subhojit
N1 - Funding Information:
We thank Sue Yeon Yi (UW-Madison) for help with constructs; and Karen Jansen-West and Lillian M Daugherty (Mayo Clinic, Jacksonville) for AAV packaging and purification. J.S. is funded by a postdoctoral fellowship from the Alzheimer’s Association (AARF-17-529623). A.A.S. is supported by The Henry and Marilyn Taub Foundation. This work was supported by NIH grants (R01AG048218 and R21 AG052404), and a “UW2020 grant” from the University of Wisconsin-Madison to S.R.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - 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.
AB - 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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U2 - 10.1038/s41467-018-07971-8
DO - 10.1038/s41467-018-07971-8
M3 - Article
C2 - 30604771
AN - SCOPUS:85059493070
SN - 2041-1723
VL - 10
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 53
ER -