{Reference Type}: Journal Article
{Title}: A novel homozygous splice-site mutation of JK gene leads to Jk(a-b-) phenotype.
{Author}: Yang J;Ni L;Li A;Li M;Ruan S;Xiang D;Zhu Z;Ye L;
{Journal}: Transfus Med
{Volume}: 34
{Issue}: 1
{Year}: 2024 Feb 10
{Factor}: 2.057
{DOI}: 10.1111/tme.13016
{Abstract}: <B>OBJECTIVE: </B>This study aimed to investigate the molecular mechanism of the Jk(a-b-) phenotype in a Chinese transfusion patient.<BR><B>BACKGROUND: </B>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.<BR><B>METHODS: </B>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.<BR><B>RESULTS: </B>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.<BR><B>CONCLUSIONS: </B>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.