Analysis of a polymorphic microRNA target site in the purinergic receptor P2RX7 gene
Analysis of a polymorphic microRNA target site in the purinergic receptor P2RX7 gene
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DOI：
10.1002/elps.200900664
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发表时间：
2010-06-01
期刊：
影响因子：
2.9
通讯作者：
Nemoda, Zsofia
中科院分区：
文献类型：
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作者：
Rahman, Omar Abdul;Sasvari-Szekely, Maria;Nemoda, Zsofia
最近对 microRNA (miRNA) 转录后调控的发现引起了我们对抑郁相关精神疾病候选基因中推定 miRNA 靶位点 SNP 的关注。 P2RX7（嘌呤能受体 P2X，配体门控离子通道，7）基因由于与 rs2230912 (G1n460Arg) 多态性重复相关，被认为是重度抑郁症和双相情感障碍的候选基因。由于该多态性位于编码区的末端，我们考虑了与 3' 非翻译区的推定 miRNA 靶位点中的 SNP 可能的连锁。根据我们的计算机搜索，rs1653625 满足此标准。然而，该 SNP 被多聚胞嘧啶和多聚腺嘌呤束包围，这阻碍了其分析至今。在这项研究中，我们描述了一种易于应用的 rs1653625 基因分型方法，通过应用引入错配核苷酸的引物来创建限制性酶切位点。通过传统的水平琼脂糖凝胶电泳和多毛细管凝胶电泳分离所得具有19个碱基对差异的等位基因特异性产物。开发的基因分型方法应用于我们的抑郁症相关关联研究。
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 (G1n460Arg) 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.