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Generation of ribosome imprinted polymers for sensitive detection of translational responses

机译:核糖体印迹聚合物的生成,用于敏感检测翻译反应

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摘要

Whilst the profiling of the transcriptome and proteome even of single-cells becomes feasible, theudanalysis of the translatome, which refers to all messenger RNAs (mRNAs) engaged with ribosomesudfor protein synthesis, is still an elaborate procedure requiring millions of cells. Herein, we report theudgeneration and use of “smart materials”, namely molecularly imprinted polymers (MIPs) to facilitateudthe isolation of ribosomes and translated mRNAs from merely 1,000 cells. In particular, we showudthat a hydrogel-based ribosome imprinted polymer could recover ribosomes and associated mRNAsudfrom human, simian and mice cellular extracts, but did not selectively enrich yeast ribosomes,udthereby demonstrating selectivity. Furthermore, ribosome imprinted polymers enabled the sensitiveudmeasurement of an mRNA translational regulatory event, requiring 1,000-fold less cells than currentudmethodologies. These results provide first evidence for the suitability of MIPs to selectively recoverudribonucleoprotein complexes such as ribosomes, founding a novel means for sensitive detection of geneudregulation.
机译:尽管转录组和蛋白质组甚至单细胞的谱图分析变得可行,但对翻译组的分析(涉及所有与核糖体结合的信使RNA(mRNA))用于蛋白质合成的ud仍然是一项复杂的程序,需要数百万个细胞。本文中,我们报道了“智能材料”的变性和使用,即分子印迹聚合物(MIP)有助于从仅1,000个细胞中分离核糖体和翻译的mRNA。特别地,我们显示印记的基于水凝胶的核糖体印迹聚合物可以从人,猿猴和小鼠细胞提取物中回收核糖体和相关的mRNA,但并未选择性地富集酵母核糖体,从而证明了其选择性。此外,核糖体印迹聚合物能够对mRNA的翻译调控事件进行灵敏的检测,比目前的方法学检测的细胞少1000倍。这些结果为MIPs选择性回收核糖核蛋白复合物(如核糖体)的适用性提供了第一个证据,为灵敏地检测基因异常调节提供了新的手段。

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