Nanotechnologies for Determining Gene Expression Patterns from Single Cells

项目摘要 所有现有的全局基因表达测定技术都需要相对大量的 分析物;为了将它们与存在于几个或单个细胞中的微量mRNA一起使用， 需要酶促扩增[5]。这个过程耗时，技术难度大， 贵了更重要的是，扩增过程本身会使样本产生偏差， 真正的量化分析[6]。这与微阵列的特定限制相结合 和DNA测序，导致没有直接的方法来研究单细胞中的转录。 为了解决这个问题，我们的目标是直接鉴定单个基因转录物分子（在基因组中）。 cDNA的形式）。本提案的具体目标是将联合收割机 硬件、软件和化学的组成部分，我们已经发展成为一个 集成的功能系统。我们提出的改进将大大影响 通过减少药物小样本转录的时间、成本和技术复杂性 分析;并将通过产生更容易解释，更好的质量， 的信息。

Project Summary All existing global gene expression assay techniques require relatively large quantities of analyte; in order to use them with the minute amount of mRNA present in a few or a single cell, enzymatic amplification is required [5]. This process is time consuming, technically difficult, and expensive. More importantly, the amplification process itself biases the sample in a way that prohibits truly quantitative analysis [6]. This, combined with specific limitations of microarrays and DNA sequencing, results in no straightforward method to study transcription in single cells. To solve this problem we aim to directly identify individual gene transcript molecules (in the form of cDNA) via atomic force microscopy. The Specific Aims of this Proposal are to combine the building blocks of hardware, software and chemistry that we have developed into an integrated, functional system. The improvement we are proposing will significantly impact medicine by reducing time, cost and technical complexity of small sample transcriptional profiling; and will lower the bioinformatics burden by producing easier to interpret, better quality of information.