Influence of SCN1A intronic mutations in mRNA splicing and relation of mRNA splicing changes with phenotype in febrile seizures related epilepsy
10.3760/cma.j.issn.1671-8925.2018.08.001
- VernacularTitle:热性惊厥相关癫痫SCN1A基因内含子突变对mRNA剪切的影响及其与临床表型的关系
- Author:
Lu YU
1
;
Heng MENG
;
Bin TANG
;
Haiqing XU
;
Xiuqu CAI
;
Na HE
;
Xiaorong LIU
;
Bingmei LI
;
Meimei GAO
;
Yiwu SHI
;
Yonghong YI
;
Weiping LIAO
Author Information
1. 广西医科大学第一附属医院神经内科
- Keywords:
SCNIA gene;
Febrile seizure related epilepsy;
Intronic mutation;
Phenotype;
In vitro splicing assay
- From:
Chinese Journal of Neuromedicine
2018;17(8):757-764
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the influence of SCNIA intronic mutations in mRNA splicing in febrile seizures related epilepsy,and investigate the association between splicing changes and genotype-phenotype-inheritance pattern.Methods Molecular cloning of 5 SCN1A intronic mutations was performed in patients with partial epilepsy with antecedent febrile seizures plus (PEFS+) and Dravet syndrome (DS) through constructing mutant and wild-type plasmids of pTragetE2-3-4-5 and E24-25-26 by using Minigene splicing assay,and the in vitro expressions in HENK293 cells were detected.The mRNA splicing changes were analyzed qualitatively and quantitatively by reverse transcription (RT)-PCR and real time quantitative (q)-PCR.Results (1) Using RT-PCR,DS mutants presented a whole exon skipping without significant remain of normal mRNA transcripts,while PEFS+ mutants showed partial exon skipping or intronic insertion with coexistence of normal and aberrant mRNA transcripts.(2) Statistical differences were found between relative quantity (RQ) of aberrant and normal mRNA in PEFS+ mutant (c.473+5G＞A:4.92％±1.05％ and 6.10％±0.21％;c.473+5G＞C:7.97％±1.12％ and 3.94％ ±1.25％) and that in DS mutant (c.602+1G＞A:60.51％±1.81％ and 0.060％±0.022％,P＜0.05);similarly,there were statistical differences between relative RQ of normal and aberrant mRNA in PEFS+ mutant c.4853-25T＞A (71.22％±11.92％ and 7.38％±1.61％) and that in DS mutant c.4853-1G＞C (0.08％±0.01％ and 22.11％±2.83％,P＜0.05).Conclusion The position and difference of splicing patterns of SCNIA intronic mutations are potential molecular pathogenesis for phenotypic difference of febrile seizures related epilepsy.
