• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>AIMS Genetics >Identification of a novel functional miR-143-5p recognition element in the Cystic Fibrosis Transmembrane Conductance Regulator 3’UTR
【24h】

Identification of a novel functional miR-143-5p recognition element in the Cystic Fibrosis Transmembrane Conductance Regulator 3’UTR

机译：囊性纤维化跨膜电导调节剂3’UTR中一种新型功能性miR-143-5p识别元件的鉴定

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

MicroRNAs (miRNAs) are small non-coding RNAs involved in regulation of gene expression. They bind in a sequence-specific manner to miRNA recognition elements (MREs) located in the 3’ untranslated region (UTR) of target mRNAs and prevent mRNA translation. MiRNA expression is dysregulated in cystic fibrosis (CF), affecting several biological processes including ion conductance in the epithelial cells of the lung. We previously reported that miR-143 is up-regulated in CF bronchial brushings compared to non-CF. Here we identified two predicted binding sites for miR-143-5p (starting at residues 558 and 644) on the CFTR mRNA, and aimed to assess whether CFTR is a true molecular target of miR-143-5p. Expression of miR-143-5p was found to be up-regulated in a panel of CF vs non-CF cell lines (1.7-fold, P = 0.0165), and its levels were increased in vitro after 20 hours treatment with bronchoalveolar lavage fluid from CF patients compared to vehicle-treated cells (3.3-fold, P = 0.0319). Luciferase assays were performed to elucidate direct miRNA::target interactions and showed that miR-143-5p significantly decreased the reporter activity when carrying the wild-type full length sequence of CFTR 3’UTR (minus 15%, P = 0.005). This repression was rescued by the disruption of the first, but not the second, predicted MRE, suggesting that the residue starting at 558 was the actual active binding site. In conclusion, we here showed that miR-143-5p modestly but significantly inhibits CFTR, improving the knowledge on functional MREs within the CFTR 3’UTR. This could lead to the development of novel therapeutic strategies where miRNA-mediated CFTR repression is blocked thereby possibly increasing the efficacy of the currently available CFTR modulators.
机译：MicroRNA（miRNA）是参与基因表达调控的小型非编码RNA。它们以序列特异性的方式与位于靶标mRNA的3'非翻译区（UTR）的miRNA识别元件（MRE）结合，并阻止mRNA的翻译。在囊性纤维化（CF）中，miRNA的表达失调，影响了几种生物学过程，包括肺上皮细胞中的离子传导。我们先前曾报道，与非CF相比，CF支气管刷中miR-143上调。在这里，我们确定了CFTR mRNA上两个预测的miR-143-5p结合位点（起始于残基558和644），旨在评估CFTR是否是miR-143-5p的真正分子靶标。发现在一组CF与非CF细胞系中，miR-143-5p的表达上调（1.7倍，P = 0.0165），并且在用支气管肺泡灌洗液处理20小时后，其水平升高与接受媒介物处理的细胞相比，来自CF患者的血样（3.3倍，P = 0.0319）。进行荧光素酶分析以阐明直接的miRNA ::靶标相互作用，结果表明当携带野生型CFTR 3'UTR全长序列时，miR-143-5p显着降低了报道分子的活性（负15％，P = 0.005）。这种抑制作用是通过破坏第一个但不是第二个预测的MRE来挽救的，这表明起始于558的残基是实际的活性结合位点。总之，我们在这里表明miR-143-5p适度但会显着抑制CFTR，从而提高了CFTR 3’UTR中功能性MRE的知识。这可能会导致开发新的治疗策略，其中miRNA介导的CFTR阻滞作用被阻断，从而可能提高当前可用CFTR调节剂的功效。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号