Functional analysis of a new splicing mutation in the <em>MYBPC3</em> gene in hypertrophic cardiomyopathy
Aim. To study the pathogenic effect in the MYBPC3 splice-site variant in the patient with hypertrophic cardiomyopathy. Materials and methods. The study was conducted using a DNA sample obtained from a patient with hypertrophic cardiomyopathy, in whom a previously undescribed variant was identified i...
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Siberian State Medical University (Tomsk)
2024-07-01
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| Series: | Бюллетень сибирской медицины |
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| Online Access: | https://bulletin.ssmu.ru/jour/article/view/5675 |
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| author | R. R. Salakhov M. V. Golubenko M. Y. Skoblov R. R. Savchenko N. R. Valiakhmetov E. N. Pavlyukova M. S. Nazarenko |
| author_facet | R. R. Salakhov M. V. Golubenko M. Y. Skoblov R. R. Savchenko N. R. Valiakhmetov E. N. Pavlyukova M. S. Nazarenko |
| author_sort | R. R. Salakhov |
| collection | DOAJ |
| description | Aim. To study the pathogenic effect in the MYBPC3 splice-site variant in the patient with hypertrophic cardiomyopathy. Materials and methods. The study was conducted using a DNA sample obtained from a patient with hypertrophic cardiomyopathy, in whom a previously undescribed variant was identified in the splice donor site of intron 21. The methods used included constructing and cloning of minigenes (vector pSpl3-Flu2-TKdel) and transfection of a human cell culture (HEK293T), followed by isolation of mRNA, production of cDNA, PCR of the minigene region containing the analyzed fragment, agarose gel electrophoresis, and Sanger sequencing. Results. The chr11:47339649-A-C (hg38) variant, disrupting the splice donor site in intron 21 (NM_000256.3: c.2067+2T>G), was identified in the 23-year-old patient with obstructive hypertrophic cardiomyopathy. To directly analyze the effect of this variant on splicing, a vector containing exon 21, intron 21, exon 22, and partially introns 20 and 22 of the MYBPC3 gene was obtained. A comparison of mRNAs from the minigenes containing / not containing the variant showed that the chr11:47339649-A-C substitution led to exon 21 and exon 22 skipping during splicing. Conclusion. The study established the functional significance of the previously undescribed variant c.2067+2T>G in the MYBPC3 gene, resulting in disruption of the mRNA splicing mechanism in the patient with hypertrophic cardiomyopathy. This variant can be classified as pathogenic. |
| format | Article |
| id | doaj-art-716e04dffe4e48f4a80b481418f33d30 |
| institution | Kabale University |
| issn | 1682-0363 1819-3684 |
| language | English |
| publishDate | 2024-07-01 |
| publisher | Siberian State Medical University (Tomsk) |
| record_format | Article |
| series | Бюллетень сибирской медицины |
| spelling | doaj-art-716e04dffe4e48f4a80b481418f33d302025-08-20T04:00:13ZengSiberian State Medical University (Tomsk)Бюллетень сибирской медицины1682-03631819-36842024-07-0123218318910.20538/1682-0363-2024-2-183-1893084Functional analysis of a new splicing mutation in the <em>MYBPC3</em> gene in hypertrophic cardiomyopathyR. R. Salakhov0M. V. Golubenko1M. Y. Skoblov2R. R. Savchenko3N. R. Valiakhmetov4E. N. Pavlyukova5M. S. Nazarenko6Research Institute of Medical Genetics, Tomsk National Research Medical Center (NRMC), Russian Academy of SciencesResearch Institute of Medical Genetics, Tomsk National Research Medical Center (NRMC), Russian Academy of SciencesResearch Center of Medical GeneticsResearch Institute of Medical Genetics, Tomsk National Research Medical Center (NRMC), Russian Academy of SciencesResearch Institute of Medical Genetics, Tomsk National Research Medical Center (NRMC), Russian Academy of SciencesCardiology Research Institute, Tomsk National Research Medical Center (NRMC), Russian Academy of SciencesResearch Institute of Medical Genetics, Tomsk National Research Medical Center (NRMC), Russian Academy of SciencesAim. To study the pathogenic effect in the MYBPC3 splice-site variant in the patient with hypertrophic cardiomyopathy. Materials and methods. The study was conducted using a DNA sample obtained from a patient with hypertrophic cardiomyopathy, in whom a previously undescribed variant was identified in the splice donor site of intron 21. The methods used included constructing and cloning of minigenes (vector pSpl3-Flu2-TKdel) and transfection of a human cell culture (HEK293T), followed by isolation of mRNA, production of cDNA, PCR of the minigene region containing the analyzed fragment, agarose gel electrophoresis, and Sanger sequencing. Results. The chr11:47339649-A-C (hg38) variant, disrupting the splice donor site in intron 21 (NM_000256.3: c.2067+2T>G), was identified in the 23-year-old patient with obstructive hypertrophic cardiomyopathy. To directly analyze the effect of this variant on splicing, a vector containing exon 21, intron 21, exon 22, and partially introns 20 and 22 of the MYBPC3 gene was obtained. A comparison of mRNAs from the minigenes containing / not containing the variant showed that the chr11:47339649-A-C substitution led to exon 21 and exon 22 skipping during splicing. Conclusion. The study established the functional significance of the previously undescribed variant c.2067+2T>G in the MYBPC3 gene, resulting in disruption of the mRNA splicing mechanism in the patient with hypertrophic cardiomyopathy. This variant can be classified as pathogenic.https://bulletin.ssmu.ru/jour/article/view/5675hypertrophic cardiomyopathymybpc3minigenesplicing |
| spellingShingle | R. R. Salakhov M. V. Golubenko M. Y. Skoblov R. R. Savchenko N. R. Valiakhmetov E. N. Pavlyukova M. S. Nazarenko Functional analysis of a new splicing mutation in the <em>MYBPC3</em> gene in hypertrophic cardiomyopathy Бюллетень сибирской медицины hypertrophic cardiomyopathy mybpc3 minigene splicing |
| title | Functional analysis of a new splicing mutation in the <em>MYBPC3</em> gene in hypertrophic cardiomyopathy |
| title_full | Functional analysis of a new splicing mutation in the <em>MYBPC3</em> gene in hypertrophic cardiomyopathy |
| title_fullStr | Functional analysis of a new splicing mutation in the <em>MYBPC3</em> gene in hypertrophic cardiomyopathy |
| title_full_unstemmed | Functional analysis of a new splicing mutation in the <em>MYBPC3</em> gene in hypertrophic cardiomyopathy |
| title_short | Functional analysis of a new splicing mutation in the <em>MYBPC3</em> gene in hypertrophic cardiomyopathy |
| title_sort | functional analysis of a new splicing mutation in the em mybpc3 em gene in hypertrophic cardiomyopathy |
| topic | hypertrophic cardiomyopathy mybpc3 minigene splicing |
| url | https://bulletin.ssmu.ru/jour/article/view/5675 |
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