Nanotechnologies for Determining Gene Expression Patterns from Single Cells

项目摘要 所有现有的全球基因表达分析技术都需要相对大量的 分析物；为了将它们与少数细胞或单个细胞中存在的微量mRNA一起使用， 需要酶促扩增[5]。这一过程耗时、技术困难，而且 很贵的。更重要的是，扩增过程本身会使样本产生偏差 禁止真正的定量分析[6]。这一点，再加上微阵列的具体限制 以及DNA测序，导致没有直接的方法来研究单细胞的转录。 为了解决这个问题，我们的目标是直接识别单个基因转录分子(在 通过原子力显微镜观察)。这项提案的具体目标是将 硬件、软件和化学的构建块，我们已经开发成 集成的、功能强大的系统。我们提议的改进将产生重大影响 通过减少小样本转录的时间、成本和技术复杂性来实现医药 并将通过生成更易于解释、质量更好的产品来减轻生物信息学的负担 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.