Engineering nanostructured probes for sensitive intracellular gene detection.

Gang Bao, Andrew Tsourkas, Philip J Santangelo
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Abstract

The ability to detect, localize, quantify and monitor the expression of specific genes in living cells in real-time will offer unprecedented opportunities for advancement in molecular biology, disease pathophysiology, drug discovery, and medical diagnostics. However, current methods for quantifying gene expression employ either selective amplification (as in PCR) or saturation binding followed by removal of the excess probes (as in microarrays and in situ hybridization) to achieve specificity. Neither approach is applicable when detecting gene transcripts within living cells. Here we review the recent development in engineering nanostructured molecular probes for gene detection in vivo, describe probe design approaches and its structure-function relations, and discuss the critical issues and challenges in performing living cell gene detection with high specificity, sensitivity and signal-to-background ratio. The underlying biological and biochemical aspects are illustrated.

用于细胞内基因敏感检测的工程纳米结构探针。
实时检测、定位、量化和监测活细胞中特定基因表达的能力,将为分子生物学、疾病病理生理学、药物发现和医学诊断领域的进步提供前所未有的机会。然而,目前定量基因表达的方法要么采用选择性扩增(如PCR),要么采用饱和结合,然后去除多余的探针(如微阵列和原位杂交),以达到特异性。这两种方法都不适用于检测活细胞内的基因转录物。本文综述了用于体内基因检测的工程纳米结构分子探针的最新进展,描述了探针的设计方法及其结构-功能关系,并讨论了实现高特异性、高灵敏度和高信本比的活细胞基因检测的关键问题和挑战。潜在的生物学和生物化学方面的说明。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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