Analysis of a polymorphic microRNA target site in the purinergic receptor P2RX7 gene.
The recent discovery of post-transcriptional regulation by microRNAs (miRNAs) drew our attention to SNPs of putative miRNA target sites in candidate genes of depression-related psychiatric disorders. The P2RX7 (purinergic receptor P2X, ligand-gated ion channel, 7) gene has been suggested as a candidate for major depressive and bipolar disorder, because of repeated associations with the rs2230912 (Gln460Arg) polymorphism. As this polymorphism is located at the end of the coding region, we considered a possible linkage with SNP(s) in putative miRNA target sites of the 3' untranslated region. Based on our in silico search, the rs1653625 fulfilled this criterion. This SNP, however, is surrounded with polycytosine and polyadenine tracts, which hindered its analysis until now. In this study, we describe a readily applicable genotyping method for rs1653625 by applying a primer that introduces mismatched nucleotides to create a restriction enzyme cleavage site. The resulting allele-specific products with 19 base pair difference were separated by both traditional horizontal agarose gel electrophoresis and multicapillary gel electrophoresis. The developed genotyping method was applied in our depression-related association study.
Rahman
Omar Abdul
OA
Institute of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary.
Sasvari-Szekely
Maria
M
Szekely
Anna
A
Faludi
Gabor
G
Guttman
Andras
A
Nemoda
Zsofia
Z
D020137:Base Pair Mismatch; D001483:Base Sequence; D016009:Chi-Square Distribution; D003001:Cloning, Molecular; D003198:Computer Simulation; D004252:DNA Mutational Analysis / Q000379:methods; D003863:Depression; D019075:Electrophoresis, Capillary; D005838:Genotype; D006801:Humans; D015810:Linkage Disequilibrium; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D018048:Receptors, Purinergic P2 / Q000235:genetics*; D058486:Receptors, Purinergic P2X7; D015203:Reproducibility of Results
D035683:MicroRNAs; C546468:P2RX7 protein, human; D018048:Receptors, Purinergic P2; D058486:Receptors, Purinergic P2X7