首页> 外文OA文献 >A novel biochemical method to identify target genes of individual microRNAs: Identification of a new Caenorhabditis elegans let-7 target
【2h】

A novel biochemical method to identify target genes of individual microRNAs: Identification of a new Caenorhabditis elegans let-7 target

机译:鉴定单个微小RNA靶基因的新型生化方法:鉴定新的秀丽隐杆线虫let-7靶标

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务,本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文,但由于OA文献来源多样且变更频繁,仍可能出现获取不到、文献不完整或与标题不符等情况,如果获取不到我们将提供退款服务。请知悉。

摘要

MicroRNAs (miRNAs) are roughly 22-nucleotide regulatory RNAs that play important roles in many developmental and physiological processes. Animal miRNAs down-regulate target genes by forming imperfect base pairs with 3′ untranslated regions (3′ UTRs) of their mRNAs. Thousands of miRNAs have been discovered in several organisms. However, the target genes of almost all of these miRNAs remain to be identified. Here, we describe a method for isolating cDNA clones of target mRNAs that form base pairs in vivo with an endogenous miRNA of interest, in which the cDNAs are synthesized from the mRNAs using the miRNA as a reverse-transcription primer. The application of this method to Caenorhabditis elegans miRNA lin-4 under test conditions yielded many clones of the known target gene lin-14 that correspond to partial sequences 5′ to lin-4 binding sites in the 3′ UTR. The method was also applied to C. elegans miRNA let-7 and a new target gene responsible for the lethal phenotype in let-7 mutants was identified. These results demonstrate that the method is a useful way to identify targets on the basis of base pairing with individual miRNAs.
机译:微小RNA(miRNA)大约是22个核苷酸的调节性RNA,在许多发育和生理过程中都起着重要作用。动物miRNA通过与mRNA的3'非翻译区(3'UTR)形成不完美的碱基对,从而下调靶基因。在几种生物中已经发现了成千上万的miRNA。但是,几乎所有这些miRNA的靶基因仍有待鉴定。在这里,我们描述了一种与目标内源性miRNA在体内形成碱基对的目标mRNA的cDNA克隆的分离方法,其中使用miRNA作为逆转录引物从mRNA合成cDNA。该方法在测试条件下对秀丽隐杆线虫miRNA lin-4的应用产生了许多已知靶基因lin-14的克隆,这些克隆对应于3'UTR中5'到lin-4结合位点的部分序列。该方法也适用于秀丽隐杆线虫miRNA let-7,并鉴定了负责let-7突变体致死表型的新靶基因。这些结果表明,该方法是基于与单个miRNA碱基配对识别靶标的有用方法。

著录项

  • 作者

    Andachi, Yoshiki;

  • 作者单位
  • 年度 2008
  • 总页数
  • 原文格式 PDF
  • 正文语种 en
  • 中图分类

相似文献

  • 外文文献
  • 中文文献
  • 专利
代理获取

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