Splicing Characterization of CLCNKB Variants in Four Patients With Type III Bartter Syndrome

Frontiers in Genetics
Chunli WangFei Zhao

Abstract

Type III Bartter syndrome (BS) is caused by loss-of-function mutations in the gene encoding basolateral chloride channel ClC-Kb (CLCNKB), and is characterized by hypokalemic metabolic alkalosis and hyperreninemic hyperaldosteronism. Here, we investigated the molecular defects in four Chinese children with clinical manifestations of Bartter syndrome. The genomic DNA of the four patients was screened for gene variations using whole-exome sequencing (WES). The candidate variants were validated by direct Sanger sequencing. Quantitative PCR (qPCR) was subsequently performed to confirm the whole CLCNK gene deletion mutation. A minigene assay and reverse transcription PCR (RT-PCR) were performed to analyze the effect of splice variants in vitro. Our patients showed early onset age with hyponatremia, hypokalemia, hypochloremia, repeated vomiting and growth retardation, suggesting Bartter syndrome. Genetic analysis revealed that all patients carried compound heterozygous or homozygous truncating variants in the CLCNKB gene. In particular, we identified a novel nonsense variant c.239G > A (p.(Trp80*)), two splice site variants (c.1053-1 G > A and c.1228-2A > G), a whole gene deletion, and a novel synonymous variant c.228A > C (p.(Arg76Ar...Continue Reading

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Citations

Jun 4, 2020·World Journal of Pediatrics : WJP·Flavia Cristina Carvalho MradAna Cristina Simões-E-Silva
Dec 1, 2020·Frontiers in Genetics·Sai WangLeping Shao

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Methods Mentioned

BETA
PCR
transfection
electrophoresis

Software Mentioned

Human Splicing Finder
Wheeler Aligner ( BWA )
Burrows
Human Finder

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