眾所周知,人類細(xì)胞可產(chǎn)生成千上萬個(gè)不同的microRNA(miRNAs),,其中miRNAs是些小片段遺傳物質(zhì),,有助于確定特定時(shí)間打開或關(guān)閉哪一個(gè)基因。miRNAs是正常細(xì)胞功能的一個(gè)重要部分,,但能促發(fā)人類疾病,,如:特定腫瘤內(nèi)的一些miRNAs被升高,則會(huì)促進(jìn)細(xì)胞存活,。為了更好地了解miRNAs如何影響健康與疾病,,研究人員首先必須知道哪一個(gè)miRNAs作用于哪一個(gè)基因,考慮到miRNAs絕對(duì)數(shù)量與單個(gè)miRNA調(diào)控成百上千個(gè)基因的事實(shí),,這是一個(gè)很大的挑戰(zhàn)?,F(xiàn)在,有一種名為miR-TRAP的新方法問世,,它使用方便,,可直接鑒定細(xì)胞內(nèi)miRNA靶標(biāo)。 與現(xiàn)有鑒定miRNA靶標(biāo)的方法相比,,miR-TRAP更簡(jiǎn)單,、更精確,主要理由有3個(gè),,分別是:首先,,miR-TRAP可直接鑒定正?;蚣膊∏闆r下活體細(xì)胞內(nèi)的miRNA靶標(biāo);其次,,不僅能識(shí)別出被miRNAs減少的mRNA靶標(biāo),,也能識(shí)別翻譯被抑制的蛋白質(zhì)靶標(biāo),這一靶標(biāo)通常不能被其他技術(shù)檢測(cè)出來,,如熒光定量PCR或芯片分析,;第三,不依賴于抗體,,因?yàn)榭贵w能導(dǎo)致非特異性背景信號(hào)和復(fù)雜數(shù)據(jù)解釋,。通過對(duì)2個(gè)重要miRNAs的13個(gè)預(yù)測(cè)靶標(biāo)的分析,這項(xiàng)技術(shù)不但確認(rèn)了已知的基因靶標(biāo),,還揭示了2個(gè)新靶標(biāo),。(生物谷bioon.com) miR-TRAP: A Benchtop Chemical Biology Strategy to Identify microRNA Targets Huricha Baigude, Ahsanullah, Zhonghan Li, Ying Zhou, Tariq M. Rana It′s a TRAP! microRNAs (miRNAs) are noncoding small RNAs that consist of approximately 21 nucleotides and are assembled into an RNA-induced silencing complex (RISC), which suppresses target mRNA expression. An easy-to-use method called miRNA target RNA affinity purification (miR-TRAP) directly identifies miRNA targets in vivo. This method could be widely used to discover miRNA targets in various disease models under physiological conditions. |
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