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However, this approach involve cell reprogramming from somatic tissue biopsies or genomic editing in healthy iPSCs for every mutation found and to be investigated. We aim to knockout (KO) Na<jats:sub>V<\/jats:sub>1.5, the cardiac sodium channel, in a healthy human iPSC line, characterize the model and then, use it to express variants of Na<jats:sub>V<\/jats:sub>1.5. We develop a homozygous Na<jats:sub>V<\/jats:sub>1.5 KO iPSC line able to differentiate into cardiomyocytes with CRISPR\/Cas9 tool. The Na<jats:sub>V<\/jats:sub>1.5 KO iPSC-CMs exhibited an organized contractile apparatus, spontaneous contractile activity, and electrophysiological recordings confirmed the major reduction in total Na<jats:sup>+<\/jats:sup> currents. The action potentials (APs) exhibited a reduction in their amplitude and in their maximal rate of rise. Voltage optical mapping recordings revealed that the conduction velocity Ca<jats:sup>2+<\/jats:sup> transient waves propagation velocities were slow. A wild-type (WT) Na<jats:sub>V<\/jats:sub>1.5 channel expressed by transient transfection in the KO iPSC-CMs restored Na<jats:sup>+<\/jats:sup> channel expression and AP properties. The expression of Na<jats:sub>V<\/jats:sub>1.5\/delQKP, a long QT type 3 (LQT3) variant, in the Na<jats:sub>V<\/jats:sub>1.5 KO iPSC-CMs showed that dysfunctional Na<jats:sup>+<\/jats:sup> channels exhibited a persistent Na<jats:sup>+<\/jats:sup> current and caused prolonged AP duration that led to arrhythmic events, characteristics of LQT3.<\/jats:p>","DOI":"10.1038\/s41598-021-96474-6","type":"journal-article","created":{"date-parts":[[2021,8,25]],"date-time":"2021-08-25T10:46:44Z","timestamp":1629888404000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["NaV1.5 knockout in iPSCs: a novel approach to study NaV1.5 variants in a human cardiomyocyte environment"],"prefix":"10.1038","volume":"11","author":[{"given":"Marion","family":"Pierre","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammed","family":"Djemai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hugo","family":"Poulin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed","family":"Chahine","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,8,25]]},"reference":[{"key":"96474_CR1","doi-asserted-by":"publisher","first-page":"H561","DOI":"10.1152\/ajpheart.1979.236.4.H561","volume":"236","author":"E Coraboeuf","year":"1979","unstructured":"Coraboeuf, E., Deroubaix, E. & Coulombe, A. 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