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

It is clinically important to detect the single-nucleotide polymorphism (SNP) of CYP3A53 (6986A>G) associated with enzymatic activity for drug metabolism. The aim of this study was to establish an accurate strategy for SNP screening.

Polyacrylamide gel electrophoresis (PAGE) using zinc(II) complexes were applied for SNP detection. Genomic analyses of 19 healthy subjects were conducted by using both Zn(2+)-Phos-tag-PAGE and Zn(2+)-cyclen-PAGE methodologies.

Zn(2+)-Phos-tag PAGE permitted identification of the following allele genotypes: the A/A homozygote (CYP3A51/1) in 3 individuals, the G/G homozygote (CYP3A53/3) in 14 individuals, and the A/G heterozygote (CYP3A51/3) in 2 individuals. Zn(2+)-cyclen PAGE demonstrated not only reproducibility of the genotyping but also existence of a novel heterozygous SNP (6929G>A) in the subject with CYP3A51/1.

We demonstrated reliable SNP genotyping and mapping in CYP3A5 using the combination method of Zn(2+)-Phos-tag PAGE and Zn(2+)-cyclen PAGE.