Screening inhibitory potential of anti-HIV RT RNA aptamers.
Screening inhibitory potential of anti-HIV RT RNA aptamers.
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DOI：
10.1007/978-1-62703-730-3_2
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发表时间：
2014-01-01
期刊：
影响因子：
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通讯作者：
Burke, Donald H
中科院分区：
文献类型：
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作者：
Lange, Margaret J;Burke, Donald H
针对 HIV 逆转录酶 (RT) 的适体已被证明可以在生化测定和细胞培养中抑制 RT。然而，迄今为止用于评估病毒抑制的方法利用了感染性HIV在表达适体的稳定细胞系中耗时的连续传代。我们建立了一种基于转染的快速检测系统，可有效检测两种催化失活锤头核酶之间表达的抗 HIV RT 适体的抑制潜力。我们的系统可以针对各种克隆方案进行更改和优化，并且向盒中添加感兴趣的序列非常简单明了。当与分析适体 RNA 在细胞中积累和定位以及包装到假型病毒粒子的方法配合使用时，该方法在预测良好的抑制剂方面具有非常高的成功率。
Aptamers targeted to HIV reverse transcriptase (RT) have been demonstrated to inhibit RT in biochemical assays and as in cell culture. However, methods employed to date to evaluate viral suppression utilize time-consuming serial passage of infectious HIV in aptamer-expressing stable cell lines. We have established a rapid, transfection-based assay system to effectively examine the inhibitory potential of anti-HIV RT aptamers expressed between two catalytically inactive hammerhead ribozymes. Our system can be altered and optimized for a variety of cloning schemes, and addition of sequences of interest to the cassette is simple and straightforward. When paired with methods to analyze aptamer RNA accumulation and localization in cells and as packaging into pseudotyped virions, the method has a very high level of success in predicting good inhibitors.