Reverse Transcription Free Microarray Analysis

描述（由申请人提供）：本项目的目标是开发一种新的微阵列系统，用于直接用总RNA进行基因表达分析，而无需标记和逆转录成cDNA。这一目标的实现将为微阵列领域提供一个关键的突破，并建立一个新的标准。事实上，尽管微阵列技术已经被广泛研究并在许多应用中被证明是成功的，但在该领域仍然存在许多挑战。首先，传统的微阵列检测需要将总RNA逆转录并标记报告分子，这是昂贵且耗时的。第二，靶序列的PCR扩增可能引起系统误差和偏倚的杂交信号。第三，微阵列仪器的成本很高。降低仪器和日常操作的成本将使小型研究机构和诊所更容易获得微阵列技术。该项目的关键创新是一种新的RNA检测技术，该技术利用目标分子和纳米颗粒的静电相互作用。该检测技术允许直接使用总RNA进行杂交实验，而无需报告基因掺入和从RNA到cDNA的逆转录。该技术还允许在基因表达分析中使用廉价的仪器来高灵敏度地检测DNA和RNA。随着最近在肿瘤发生和化疗结果预后的临床测试中使用微阵列的报道（Science，303：1754），仅在美国，引入先进且廉价的微阵列平台将在500多个临床实验室和数百个生命科学研究实验室中快速采用。

DESCRIPTION (provided by applicant): The goal of this project is to develop a new microarray system for performing gene expression analysis directly with total RNA without labeling and reverse transcription to cDNA. Achieving this goal will provide a critical breakthrough and establish a new standard in microarray field. Indeed, although microarray technique has been widely studied and proved to be successful in many applications, there are still many challenges in this field. First, the traditional microarray detection requires the total RNA to be reverse transcribed and labeled with reporter molecules, which is costly and time-consuming. Second, the PCR amplification of target sequences may give rise to systematic errors and biased hybridization signals. Third, the cost for a microarray instrumentation is high. Reducing the cost of instrumentation and day-by-day operation will make microarray technology more accessible to small research institutions and clinics. The key innovation of this project is a new RNA detection technique which employs electrostatic interaction of the target molecules and nanometer-size particles. This detection technique allows to use total RNA directly for hybridization experiment without reporter incorporation and reverse transcription from RNA to cDNA. The technique also allows inexpensive instrumentation to be used for highly-sensitive detection of DNA and RNA in gene expression analysis. With the recent reports of using microarrays in clinical tests for tumorogenesis and prognosis of a chemotherapy outcome (Science, 303:1754), introducing an advanced and inexpensive microarray platform will find fast adoption in more than 500 clinical laboratories and hundreds of life science research laboratories in US alone.