RESUMO
Loss-of-function (LoF) mutations in FLNC are strongly associated with dilated cardiomyopathy (DCM). Using CRISPR/Cas9 mediated edition in an healthy donor derived iPSC (ICAN-403.3) we subcloned 1 iPSC line harboring LoF mutation in FLNC. All lines are fully pluripotent and isogenic except at edited site where it presents a homozygous (ICAN-FLNC42.1) deletion of splice site leading to skipping of exon 42 traduced into a short filamin form with reduced expression in derived cardiomyocytes. This line would serve for FLNC mutation DCM modeling after differentiation into cardiocytes or beating organoids.
Assuntos
Cardiomiopatia Dilatada , Células-Tronco Pluripotentes Induzidas , Sistemas CRISPR-Cas/genética , Cardiomiopatia Dilatada/genética , Éxons/genética , Filaminas/genética , Filaminas/metabolismo , Humanos , Células-Tronco Pluripotentes Induzidas/metabolismo , MutaçãoRESUMO
MYH7 is a major gene responsible for hypertrophic cardiomyopathy (HCM). From patient's skin fibroblasts, we derived an iPSC line (CDGEN1.16) harboring the heterozygous MYH7 R403L mutation, a hot-spot codon in HCM. We subsequently corrected the mutated codon using CRISPR/Cas9 editing and obtained the isogenic control line (CDGEN1.16.40.5) preserving the genomic background of the patient. Both lines were pluripotent and could be efficiently committed to beating cardiomyocytes (CM) suitable for subsequent cell or pseudo-tissue study of HCM pathology.
Assuntos
Cardiomiopatia Hipertrófica , Células-Tronco Pluripotentes Induzidas , Sistemas CRISPR-Cas/genética , Miosinas Cardíacas/genética , Cardiomiopatia Hipertrófica/genética , Humanos , Mutação , Miócitos Cardíacos , Cadeias Pesadas de Miosina/genéticaRESUMO
Human pluripotent stem cells offer a limitless source of cells for regenerative medicine. They are also highly valuable as tools to study development and pathologies or as cellular substrates to screen and test new drugs. We generated human induced pluripotent stem cell (hiPSC) lines from two unrelated healthy control donors. Peripheral blood mononuclear cells (PBMCs) from these donors were reprogrammed by non-integrative viral transduction, had normal karyotypes and expressed pluripotency hallmarks.