Hereditary spherocytosis due to a novel c. 5798+ 1G>A variant of the SPTB gene
10.3760/cma.j.issn.1003-9406.2020.01.005
- VernacularTitle: SPTB基因c.5798＋1G>A新变异导致遗传性球形红细胞增多症一家系
- Author:
Hongyan LIU
1
;
Jia HUANG
;
Yinghai JIANG
;
Liangjie GUO
;
Hai XIAO
;
Hongdan WANG
Author Information
1. Institute of Medical Genetics, Henan Provincial People’s Hospital of Zhengzhou University, People’s Hospital of Henan University, Zhengzhou, Henan 450003, China
- Publication Type:Journal Article
- Keywords:
SPTB gene;
Hereditary spherocytosis;
Splice site;
Gene variant;
Next generation sequencing
- From:
Chinese Journal of Medical Genetics
2020;37(1):17-20
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the genetic basis of a pedigree affected with hereditary spherocytosis.
Methods:Peripheral blood samples were collected from 17 members of the pedigree. Genomic DNA of the proband was subjected to next generation sequencing. Candidate variant was validated by co-segregation analysis. pCAS2c.5798+ 1G and pCAS2c.5798+ 1A plasmids were constructed by homologous recombination and transfected into 293T cells. Reverse transcription PCR, TA cloning and Sanger sequencing were used to analyze the effect of candidate variant on splicing. Meanwhile, peripheral blood RNAs were extracted to analyze the effect of candidate variant on splicing in vivo.
Results:The proband was found to carry a c. 5798+ 1G>A variant of the SPTB gene. The variant has co-segregated with the phenotype in the pedigree. In vitro and in vivo splicing experiments confirmed that the mutation has significantly affected the splicing, resulting in shift of reading frame and produced a premature termination codon.
Conclusion:The novel c. 5798+ 1G>A variant of the SPTB gene probably underlies the pathogenesis of hereditary spherocytosis in this pedigree.
