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CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons
Basal forebrain cholinergic neurons (BFCNs) are believed to be one of the first cell types to be affected in all forms of AD, and their dysfunction is clinically correlated with impaired short-term memory formation and retrieval. We present an optimized in vitro protocol to generate human BFCNs from...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5660456/ https://www.ncbi.nlm.nih.gov/pubmed/29078805 http://dx.doi.org/10.1186/s40478-017-0475-z |
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author | Ortiz-Virumbrales, Maitane Moreno, Cesar L. Kruglikov, Ilya Marazuela, Paula Sproul, Andrew Jacob, Samson Zimmer, Matthew Paull, Daniel Zhang, Bin Schadt, Eric E. Ehrlich, Michelle E. Tanzi, Rudolph E. Arancio, Ottavio Noggle, Scott Gandy, Sam |
author_facet | Ortiz-Virumbrales, Maitane Moreno, Cesar L. Kruglikov, Ilya Marazuela, Paula Sproul, Andrew Jacob, Samson Zimmer, Matthew Paull, Daniel Zhang, Bin Schadt, Eric E. Ehrlich, Michelle E. Tanzi, Rudolph E. Arancio, Ottavio Noggle, Scott Gandy, Sam |
author_sort | Ortiz-Virumbrales, Maitane |
collection | PubMed |
description | Basal forebrain cholinergic neurons (BFCNs) are believed to be one of the first cell types to be affected in all forms of AD, and their dysfunction is clinically correlated with impaired short-term memory formation and retrieval. We present an optimized in vitro protocol to generate human BFCNs from iPSCs, using cell lines from presenilin 2 (PSEN2) mutation carriers and controls. As expected, cell lines harboring the PSEN2 (N141I) mutation displayed an increase in the Aβ42/40 in iPSC-derived BFCNs. Neurons derived from PSEN2 (N141I) lines generated fewer maximum number of spikes in response to a square depolarizing current injection. The height of the first action potential at rheobase current injection was also significantly decreased in PSEN2 (N141I) BFCNs. CRISPR/Cas9 correction of the PSEN2 point mutation abolished the electrophysiological deficit, restoring both the maximal number of spikes and spike height to the levels recorded in controls. Increased Aβ42/40 was also normalized following CRISPR/Cas-mediated correction of the PSEN2 (N141I) mutation. The genome editing data confirms the robust consistency of mutation-related changes in Aβ42/40 ratio while also showing a PSEN2-mutation-related alteration in electrophysiology. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article doi: (10.1186/s40478-017-0475-z) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5660456 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-56604562017-10-31 CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons Ortiz-Virumbrales, Maitane Moreno, Cesar L. Kruglikov, Ilya Marazuela, Paula Sproul, Andrew Jacob, Samson Zimmer, Matthew Paull, Daniel Zhang, Bin Schadt, Eric E. Ehrlich, Michelle E. Tanzi, Rudolph E. Arancio, Ottavio Noggle, Scott Gandy, Sam Acta Neuropathol Commun Research Basal forebrain cholinergic neurons (BFCNs) are believed to be one of the first cell types to be affected in all forms of AD, and their dysfunction is clinically correlated with impaired short-term memory formation and retrieval. We present an optimized in vitro protocol to generate human BFCNs from iPSCs, using cell lines from presenilin 2 (PSEN2) mutation carriers and controls. As expected, cell lines harboring the PSEN2 (N141I) mutation displayed an increase in the Aβ42/40 in iPSC-derived BFCNs. Neurons derived from PSEN2 (N141I) lines generated fewer maximum number of spikes in response to a square depolarizing current injection. The height of the first action potential at rheobase current injection was also significantly decreased in PSEN2 (N141I) BFCNs. CRISPR/Cas9 correction of the PSEN2 point mutation abolished the electrophysiological deficit, restoring both the maximal number of spikes and spike height to the levels recorded in controls. Increased Aβ42/40 was also normalized following CRISPR/Cas-mediated correction of the PSEN2 (N141I) mutation. The genome editing data confirms the robust consistency of mutation-related changes in Aβ42/40 ratio while also showing a PSEN2-mutation-related alteration in electrophysiology. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article doi: (10.1186/s40478-017-0475-z) contains supplementary material, which is available to authorized users. BioMed Central 2017-10-27 /pmc/articles/PMC5660456/ /pubmed/29078805 http://dx.doi.org/10.1186/s40478-017-0475-z Text en © The Author(s). 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Ortiz-Virumbrales, Maitane Moreno, Cesar L. Kruglikov, Ilya Marazuela, Paula Sproul, Andrew Jacob, Samson Zimmer, Matthew Paull, Daniel Zhang, Bin Schadt, Eric E. Ehrlich, Michelle E. Tanzi, Rudolph E. Arancio, Ottavio Noggle, Scott Gandy, Sam CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons |
title | CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons |
title_full | CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons |
title_fullStr | CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons |
title_full_unstemmed | CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons |
title_short | CRISPR/Cas9-Correctable mutation-related molecular and physiological phenotypes in iPSC-derived Alzheimer’s PSEN2(N141I) neurons |
title_sort | crispr/cas9-correctable mutation-related molecular and physiological phenotypes in ipsc-derived alzheimer’s psen2(n141i) neurons |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5660456/ https://www.ncbi.nlm.nih.gov/pubmed/29078805 http://dx.doi.org/10.1186/s40478-017-0475-z |
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