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Author:
Date:
2020-08-20
[Abstract] The nucleotides involved in RNA-RNA interaction can be tagged by chemical- or UV-induced crosslinking, and further identified by classical or modern high throughput techniques. The contacts of mRNA with 18S rRNA that occur along the mRNA channel of 40S subunit have been mapped by site-specific UV crosslinking followed by reverse transcriptase termination sites (RTTS) using radioactive or fluorescent oligonucleotides. However, the sensitivity of this technique is restricted to the detection of those fragments that resulted from the most frequent crosslinkings. Here, we combined RTTS with RNAseq to map the mRNA-18S rRNA contacts with a much deeper resolution. Although aimed to detect the interaction of mRNA with the ES6S region of 18S rRNA, this technique can also be applied to map the ...
[摘要] [摘要 ] 可以通过化学或紫外线诱导的交联来标记参与RNA-RNA相互作用的核苷酸，并通过经典或现代的高通量技术对其进行进一步鉴定。沿着40S亚基的mRNA通道发生的18S rRNA与mRNA的接触已通过位点特异性UV交联进行了定位，随后使用放射性或荧光寡核苷酸进行了逆转录酶终止位点（RTTS）。但是，该技术的敏感性仅限于检测由最频繁的交联产生的那些片段。在这里，我们将RTTS与RNAseq结合使用，以更深的分辨率绘制了mRNA-18S rRNA接触图。尽管旨在检测mRNA与18S rRNA的ES6S区域的相互作用，但该技术也可以用于绘制mRNA与其他非编码RNA分子的相互作用（在转录，剪接或RNA介导的转录后调控过程中（例如，snRNA，microRNA和lncRNA）。
[背景 ] 是与非编码RNA的mRNA的相互作用volved mRNA中的生命周期的每个步骤，从它的生物合成和处理进入细胞核至细胞质中的翻译和最终降解。这些相互作用可以在大分子机器核糖体和剪接体，以及在较小的复合物如RISC或者发生（RNA诱导的沉默复合物）或lncRNA介导的基因表达调节期间（皮萨列夫等人，2008; Engreitz 。等人，2014 Sharma ...
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