[因SPTB基因新型c.5798+1G>A变异导致的遗传性球形红细胞增多症]
[Hereditary spherocytosis due to a novel c.5798+1G>A variant of the SPTB gene].
作者信息
Liu Hongyan, Huang Jia, Jiang Yinghai, Guo Liangjie, Xiao Hai, Wang Hongdan
机构信息
Institute of Medical Genetics, Henan Provincial People's Hospital of Zhengzhou University, People's Hospital of Henan University, Zhengzhou, Henan 450003, China.
出版信息
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2020 Jan 10;37(1):17-20. doi: 10.3760/cma.j.issn.1003-9406.2020.01.005.
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. pCAS2(c.5798+1G) and pCAS2(c.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.
目的
探究一个患有遗传性球形红细胞增多症家系的遗传基础。
方法
采集该家系17名成员的外周血样本。对先证者的基因组DNA进行二代测序。通过共分离分析验证候选变异。采用同源重组构建pCAS2(c.5798+1G)和pCAS2(c.5798+1A)质粒，并转染至293T细胞。利用逆转录PCR、TA克隆和桑格测序分析候选变异对剪接的影响。同时，提取外周血RNA分析候选变异在体内对剪接的影响。
结果
发现先证者携带SPTB基因c.5798+1G>A变异。该变异在家系中与表型共分离。体外和体内剪接实验证实该突变显著影响剪接，导致阅读框移位并产生提前终止密码子。
结论
SPTB基因新的c.5798+1G>A变异可能是该家系遗传性球形红细胞增多症发病机制的基础。
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