Generation of Induced Pluripotent Stem Cell Lines ICGi022-A-3, ICGi022-A-4, and ICGi022-A-5 with p.Asn515del Mutation Introduced in  MYBPC3  Using CRISPR/Cas9

S. V. Pavlova; Павлова С. В.; L. Sh. Shayakhmetova; Шаяхметова Л. Ш.; K. A. Pronyaeva; Проняева К. А.; A. E. Shulgina; Шульгина А. Е.; S. M. Zakian; Закиян С. М.; E. V. Dementyeva; Дементьева Е. В.

doi:10.31857/S0475145023010111

Generation of Induced Pluripotent Stem Cell Lines ICGi022-A-3, ICGi022-A-4, and ICGi022-A-5 with p.Asn515del Mutation Introduced in MYBPC3 Using CRISPR/Cas9

Авторлар: Pavlova S.V.¹, Shayakhmetova L.S.¹, Pronyaeva K.A.¹, Shulgina A.E.¹, Zakian S.M.¹, Dementyeva E.V.¹
Мекемелер:
1. Federal Research Center Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences
Шығарылым: Том 54, № 1 (2023)
Беттер: 105-113
Бөлім: Collection of pluripotent stem cell lines
URL: https://journals.rcsi.science/0475-1450/article/view/137048
DOI: https://doi.org/10.31857/S0475145023010111
EDN: https://elibrary.ru/FRUOIN
ID: 137048

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Аннотация

Up to 60% of hypertrophic cardiomyopathy cases are due to mutations responsible for sarcomere functioning. However, clinical significance is known not for all variants found in hypertrophic cardiomyopathy-associated genes. Using methods of nucleotide sequence editing opens new prospects for clarifying clinical significance of the variants. A trinucleotide deletion c.1543_1545delAAC (p.Asn515del) with uncertain significance was introduced in MYBPC3 of induced pluripotent stem cells (iPSCs) of a healthy donor by CRISPR/Cas9. Three iPSC lines (ICGi022-A-3, ICGi022-A-4, and ICGi022-A-5) homozygous at the mutation were generated. The iPSC lines with the introduced deletion demonstrated morphology characteristic of human pluripotent cells, normal karyotype (46,XX), expressed markers of the pluripotent state (OCT4, NANOG, TRA-1-60, SSEA-4), and were able to give rise to derivatives of three germ layers during spontaneous differentiation. Studying properties of cardiomyocytes obtained under directed differentiation of the ICGi022-A-3, ICGi022-A-4, and ICGi022-A-5 iPSC lines allows establishing pathogenetic contribution of the p.Asn515del variant in MYBPC3 to hypertrophic cardiomyopathy development.

Негізгі сөздер

hypertrophic cardiomyopathy, variants of unknown significance, induced pluripotent stem cells, CRISPR/Cas9

Әдебиет тізімі

Дементьева Е.В., Вяткин Ю.В., Кретов Е.И. и др. Генетический анализ пациентов с гипертрофической кардиомиопатией // Гены & Клетки. 2020. Т. XV. № 3. С. 68–73.
Поляк М.Е., Ховалыг А.Б., Букаева А.А. и др. Спектр мутаций в гене MYBPC3 у пациентов с гипертрофической кардиомиопатией // Медицинская генетика. 2016. Т. 15. № 8. С. 26–29.
Dementyeva E.V., Pavlova S.V., Chernyavsky A.M., Zakian S.M. Generation of an induced pluripotent stem cell line, ICGi029-A, by reprogramming peripheral blood mononuclear cells of a patient suffering from hypertrophic cardiomyopathy and carrying a heterozygous p.N515del mutation in MYBPC3 // Stem Cell Res. 2021. V. 53. 102344.
Malakhova A.A., Grigor’eva E.V., Pavlova S.V. et al. Generation of induced pluripotent stem cell lines ICGi021-A and ICGi022-A from peripheral blood mononuclear cells of two healthy individuals from Siberian population // Stem Cell Res. 2020. V. 48. 101952.
Waldmüller S., Erdmann J., Binner P. et al. Novel correlations between the genotype and the phenotype of hypertrophic and dilated cardiomyopathy: results from the German Competence Network Heart Failure // Eur. J. Heart Fail. 2011. V. 13. № 11. P. 1185–1192.