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Abstract：
FRAXopathies are caused by the expansion of the CGG repeat in the 5\'UTR of the FMR1 gene, which encodes the protein responsible for the synthesis of FMRP. This mutation leads to dramatic changes in FMRP expression at both the mRNA and protein levels. Evidence is emerging that changes in FMR1 mRNA expression can lead to the dysregulation of the miRNAs that target its 3\'UTR. In the present work, B-lymphocyte cell lines obtained from patients with FRAXopathies were used, and a wide variety of FMR1 gene activities were observed, allowing the identification of the relationships between FMR1 dysregulation and miRNA activity. We studied the expression levels of eight miRNAs that target the FMR1 gene. To prove the interaction of the studied miRNAs with FMR1, a plasmid was constructed that possesses three primary structures: the miRNA gene, with expression driven by an inducible promoter; a constitutively expressed FusionRed reporter; and an eGFP reporter followed by the 3\'UTR of the FMR1 gene. We evaluated changes in miRNA expression in response to alterations in FMR1 gene activity in a model cell line as well as interactions with some miRNAs with the FMR1 3\'UTR.
摘要：
FRAX病是由CGG重复序列在FMR1基因的5'UTR中的扩增引起的,它编码负责FMRP合成的蛋白质。这种突变导致mRNA和蛋白质水平上的FMRP表达发生巨大变化。有证据表明，FMR1mRNA表达的变化可能导致靶向其3'UTR的miRNA失调。在目前的工作中,使用从FRAX病患者获得的B淋巴细胞细胞系，并观察到各种各样的FMR1基因活性，允许鉴定FMR1失调和miRNA活性之间的关系。我们研究了靶向FMR1基因的8种miRNA的表达水平。为了证明所研究的miRNA与FMR1的相互作用，构建了具有三个一级结构的质粒：miRNA基因，表达由诱导型启动子驱动；组成型表达FusionRed报道分子；eGFP报道分子，随后是FMR1基因的3UTR。我们评估了miRNA表达的变化，以响应模型细胞系中FMR1基因活性的变化，以及与某些miRNA与FMR13'UTR的相互作用。
