A novel homozygous splice-site mutation of JK gene leads to Jk(a-b-) phenotype.
Transfus Med
; 34(1): 39-45, 2024 Feb.
Article
em En
| MEDLINE
| ID: mdl-37950522
ABSTRACT
OBJECTIVES:
This study aimed to investigate the molecular mechanism of the Jk(a-b-) phenotype in a Chinese transfusion patient.BACKGROUND:
Many different mutation types relating to Jk(a-b-) phenotype have been reported. However, the splice-site mutation is relatively rare and the related functional verification is lacking. MATERIALS ANDMETHODS:
In this study, the blood sample was collected from a transfusion patient with the Jk(a-b-) phenotype. Serotyping was performed using routine serological methods. The exons sequences and coding regions of the JK gene were amplified using polymerase chain reaction and directly sequenced. To perform a minigene splicing assay, the intronic mutation sequences were cloned into a pSPL3 splice reporting vector. The splicing reporter minigene assay was performed in HEK 293T cells.RESULTS:
The Jk(a-b-) phenotype of the blood sample was identified through serological testing. Sequencing results revealed that the sample had a novel homozygous splice-site mutation JK*02N (NM_015865.7 c.663+3A>C). Further analysis, including cDNA sequencing and minigene splicing assay, confirmed that the novel splice-site mutation resulted in exon skipping. Interestingly, different numbers of exons being skipped were obtained by the two methods.CONCLUSION:
This study revealed a novel homozygous splicing-site mutation associated with the Jk(a-b-) phenotype in Chinese population. Our results emphasise the importance of the in vitro functional method minigene splicing assay, while also acknowledging its potential limitations when compared to cDNA sequencing.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Splicing de RNA
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article