Genotyping and mapping assay of single-nucleotide polymorphisms in CYP3A5 using DNA-binding zinc(II) complexes

Objective:It is clinically important to detect the single-nucleotide polymorphism (SNP) of CYP3A5⁎3 (6986A>G) associated with enzymatic activity for drug metabolism. The aim of this study was to establish an accurate strategy for SNP screening.Design and methods:Polyacrylamide gel electrophoresis (PAGE) using zinc(II) complexes were applied for SNP detection. Genomic analyses of 19 healthy subjects were conducted by using both Zn2+–Phos-tag-PAGE and Zn2+–cyclen-PAGE methodologies.Results:Zn2+–Phos-tag PAGE permitted identification of the following allele genotypes: the A/A homozygote (CYP3A5⁎1/⁎1) in 3 individuals, the G/G homozygote (CYP3A5⁎3/⁎3) in 14 individuals, and the A/G heterozygote (CYP3A5⁎1/⁎3) in 2 individuals. Zn2+–cyclen PAGE demonstrated not only reproducibility of the genotyping but also existence of a novel heterozygous SNP (6929G>A) in the subject with CYP3A5⁎1/⁎1.Conclusion:We demonstrated reliable SNP genotyping and mapping in CYP3A5 using the combination method of Zn2+–Phos-tag PAGE and Zn2+–cyclen PAGE.