基于溶胶-凝胶的微流体系统提高了RNA适体选择的效率。

Oligonucleotides Pub Date : 2011-03-01 Epub Date: 2011-03-17 DOI:10.1089/oli.2010.0263
Ji-Young Ahn, Minjoung Jo, Pooja Dua, Dong-Ki Lee, Soyoun Kim
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引用次数: 30

摘要

RNA和DNA适体以高特异性和亲和力结合靶分子,已成为诊断和治疗研究的热点。这些适体是通过SELEX获得的,通常需要多轮选择和扩增。最近,我们已经展示了利用微流控芯片有效结合和洗脱RNA适体对靶蛋白的作用,微流控芯片包含了5个溶胶-凝胶结合液滴,其中嵌入了特定的靶蛋白。在SELEX实验中,我们证明了我们的微流控芯片极大地提高了RNA适配体对tata结合蛋白的选择效率,将产生高亲和力适配体所需的选择周期减少了约80%。许多适体与以前通过常规过滤结合SELEX分离的适体相同或同源。微流控芯片SELEX很容易扩展使用溶胶-凝胶微阵列为基础的目标复用。此外,我们表明,溶胶-凝胶嵌入的蛋白质阵列可以作为一种高通量分析,用于量化适配体的结合亲和力。
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A sol-gel-based microfluidics system enhances the efficiency of RNA aptamer selection.

RNA and DNA aptamers that bind to target molecules with high specificity and affinity have been a focus of diagnostics and therapeutic research. These aptamers are obtained by SELEX often requiring many rounds of selection and amplification. Recently, we have shown the efficient binding and elution of RNA aptamers against target proteins using a microfluidic chip that incorporates 5 sol-gel binding droplets within which specific target proteins are imbedded. Here, we demonstrate that our microfluidic chip in a SELEX experiment greatly improved selection efficiency of RNA aptamers to TATA-binding protein, reducing the number of selection cycles needed to produce high affinity aptamers by about 80%. Many aptamers were identical or homologous to those isolated previously by conventional filter-binding SELEX. The microfluidic chip SELEX is readily scalable using a sol-gel microarray-based target multiplexing. Additionally, we show that sol-gel embedded protein arrays can be used as a high-throughput assay for quantifying binding affinities of aptamers.

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来源期刊
Oligonucleotides
Oligonucleotides 生物-生化与分子生物学
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