1. Haploinsufficiency of A20 with a novel mutation of deletion of exons 2-3 of
- Author
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Maho, Shimizu, Tadashi, Matsubayashi, Hidenori, Ohnishi, Mina, Nakama, Kazushi, Izawa, Yoshitaka, Honda, and Ryuta, Nishikomori
- Subjects
Male ,Heterozygote ,Loss of Function Mutation ,Humans ,Exons ,Haploinsufficiency ,Gene Deletion ,Tumor Necrosis Factor alpha-Induced Protein 3 ,Autoimmune Diseases - Abstract
Haploinsufficiency of A20 (HA20) due to loss-of-function mutations ofGenomic DNA was extracted from the peripheral blood of the patient clinically suspected of HA20. We examined autoinflammatory disease-causing genes and performed a multiplex ligation-dependent probe amplification (MLPA) assay for copy number analysis. Next, to determine the disconnection point, genomic DNA was amplified with long-range PCR and sequenced. Finally, western blotting was carried out to measure A20 protein expression in mitogen phytohaemagglutinin (PHA)-induced T-cell blasts from the patient and a healthy volunteer.Targeted next-generation sequencing found no pathogenic mutation, but copy number variation (CNV) analysis suggested a heterozygous deletion of exons 2-3. The MLPA assay and long-range PCR confirmed the mutation. Western blotting analysis indicated a marked decrease in expression of A20 protein from the patient compared to a normal control. The results showed that this deletion was a pathogenic mutation.This study demonstrates a novel mutation resulting in a deletion of exons 2-3 of
- Published
- 2020