Generation of an NCS1 gene knockout human induced pluripotent stem cell line using CRISPR/Cas9

Smilla K Maierhof; Christian Schinke; Janine Cernoch; Lois Hew; Laura Pauline Bruske; Valeria Fernandez Vallone; Kristin Fischer; Harald Stachelscheid; Petra Huehnchen; Matthias Endres; Sebastian Diecke; Narasimha Swamy Telugu; Wolfgang Boehmerle

doi:10.1016/j.scr.2023.103253

Generation of an NCS1 gene knockout human induced pluripotent stem cell line using CRISPR/Cas9

Stem Cell Res. 2023 Dec:73:103253. doi: 10.1016/j.scr.2023.103253. Epub 2023 Nov 14.

Affiliations

¹ Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Klinik und Hochschulambulanz für Neurologie, Berlin, Germany; Berlin Institute of Health at Charité, Universitätsmedizin Berlin, Anna-Louisa-Karsch Straße 2, 10178 Berlin, Germany; Einstein Center for Neurosciences at Charité, Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany. Electronic address: smilla.maierhof@charite.de.
² Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Klinik und Hochschulambulanz für Neurologie, Berlin, Germany; Berlin Institute of Health at Charité, Universitätsmedizin Berlin, Anna-Louisa-Karsch Straße 2, 10178 Berlin, Germany.
³ Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Core Unit Pluripotent Stem Cells and Organoids (CUSCO), Charitéplatz 1, 10117 Berlin, Germany.
⁴ Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Klinik und Hochschulambulanz für Neurologie, Berlin, Germany; Einstein Center for Neurosciences at Charité, Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany.
⁵ Berlin University of Applied Sciences and Technology (BHT), Luxemburger Straße 10, 13353 Berlin, Germany.
⁶ Berlin Institute of Health at Charité - Universitätsmedizin Berlin, Core Unit Pluripotent Stem Cells and Organoids (CUSCO), Charitéplatz 1, 10117 Berlin, Germany; Berlin Institute of Health (BIH) at Charité - Universitätsmedizin Berlin, BIH Center for Regenerative Therapies (BCRT), Charitéplatz 1, 10117 Berlin, Germany.
⁷ Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Klinik und Hochschulambulanz für Neurologie, Berlin, Germany; Berlin Institute of Health at Charité, Universitätsmedizin Berlin, Anna-Louisa-Karsch Straße 2, 10178 Berlin, Germany; Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, NeuroCure Cluster of Excellence, Charitéplatz 1, 10117 Berlin, Germany.
⁸ Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt Universität zu Berlin, Klinik und Hochschulambulanz für Neurologie, Berlin, Germany; Berlin Institute of Health at Charité, Universitätsmedizin Berlin, Anna-Louisa-Karsch Straße 2, 10178 Berlin, Germany; Einstein Center for Neurosciences at Charité, Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany; Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, NeuroCure Cluster of Excellence, Charitéplatz 1, 10117 Berlin, Germany; Center for Stroke Research, Charité -Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Charitéplatz 1, 10117 Berlin, Germany; German Center for Neurodegenerative Diseases (DZNE), partner site Berlin, Germany; German Center for Cardiovascular Research (DZHK), partner site Berlin, Germany.
⁹ Technology Platform Pluripotent Stemcell, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 13125 Berlin, Germany; DZHK (German Centre for Cardiovascular Research), partner site Berlin, Germany.
¹⁰ Technology Platform Pluripotent Stemcell, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), 13125 Berlin, Germany.

PMID: 37984032
DOI: 10.1016/j.scr.2023.103253

Abstract

NCS1 (Neuronal calcium sensor protein 1) encodes a highly conserved calcium binding protein abundantly expressed in neurons. It modulates intracellular calcium homeostasis, calcium-dependent signaling pathways as well as neuronal transmission and plasticity. Here, we generated a NCS1 knockout human induced pluripotent stem cell (hiPSC) line using CRISPR-Cas9 genome editing. It shows regular expression of pluripotent markers, normal iPSC morphology and karyotype as well as no detectable off-target effects on top 6 potentially affected genes. This newly generated cell line constitutes a valuable tool for studying the role of NCS1 in the pathophysiology of various neuropsychiatric disorders and non-neurological disease.

MeSH terms

CRISPR-Cas Systems* / genetics
Calcium / metabolism
Gene Editing
Gene Knockout Techniques
Humans
Induced Pluripotent Stem Cells* / metabolism

Substances

Calcium