Functional analysis of haplotypes and promoter activity at the 5′ region of the human GABRB3 gene and associations with schizophrenia

BACKGROUND: This study investigated the effects of haplotypes T‐G and C‐A derived from NG_012836.1:g.4160T>C and NG_012836.1:g.4326G>A on protein expression levels in vitro and identified the functional sequence in the regulatory region of the GABRB3 gene linked to possible associations with schizophrenia. METHODS: Recombinant plasmids with haplotypes T‐G and C‐A and 10 recombinant vectors containing deletion fragments from the GABRB3 gene 5′ regulatory region were transfected into HEK‐293, SK‐N‐SH, and SH‐SY5Y cells. The relative fluorescence intensity of the two haplotypes and different sequences was compared using a dual luciferase reporter assay system. RESULTS: The relative fluorescence intensity of haplotype C‐A was significantly lower than that of T‐G. We shortened the core promoter sequence of the GABRB3 gene 5′ regulation region from −177 bp to −18 bp (ATG+1). We also found an expression suppression region from −1,735 bp to −1,638 bp and an enhanced regulatory region from −1,638 bp to −1,335 bp. Multiple inhibitory functional elements were identified in the region from −680 bp to −177 bp. CONCLUSION: We demonstrated that haplotype C‐A might increase the risk of schizophrenia and found multiple regulatory regions that had an effect on GABRB3 receptor expression.