Nanotechnologies for Determining Gene Expression Patterns from Single Cells

用于确定单细胞基因表达模式的纳米技术

• 批准号: 8657227
• 负责人: Jason C Reed
• 金额: $ 32.52万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8657227
• 关键词: Nanotechnologies; Determining; Gene; Expression; Patterns

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.

项目概要 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；为了将它们与少数或单个细胞中存在的微量 mRNA 一起使用， 需要酶促扩增[5]。这个过程非常耗时，技术难度大， 昂贵的。更重要的是，扩增过程本身会对样本产生偏差， 禁止真正的定量分析[6]。这与微阵列的具体局限性相结合 和 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 cDNA 形式）通过原子力显微镜。该提案的具体目标是将 我们已将硬件、软件和化学的构建模块开发成 集成的、功能性的系统。我们提出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医学 分析；并将通过产生更容易解释、更好的质量来减轻生物信息学负担 的信息。