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

基本信息

批准号：
8146147
负责人：
Jason C Reed
金额：
$ 33.51万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
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/8146147
关键词：
Address Algorithms Applications Grants Atomic Force Microscopy Behavior Benchmarking Biochemical Biochemistry Bioinformatics Biological Biological Assay Biological Process Biology Brain California Cells Chemistry Code Collaborations Complementary DNA Complex Computer software DNA Sequence Data Analyses Databases Detection Development Digestion Disease Engineering Enzymes Experimental Designs Funding Gene Expression Genes Genetic Transcription Genomics Goals Health Hour Human Image In Situ Individual Institutes Journals Location Malignant Neoplasms Measures Medicine Messenger RNA Methods Molecular Molecular Analysis Nanotechnology New York Pattern Polynucleotides Problem Solving Procedures Process Publications Quality Control Reproducibility Research Research Project Grants Sampling Sampling Biases Scanning Probe Microscopes Scanning Probe Microscopy Science Software Tools Sorting - Cell Movement Surface System Systems Biology Techniques Testing Time Tissues Transcript United States National Institutes of Health Universities Vertebral column Work base cost design digital image processing instrument interdisciplinary collaboration mathematical model member nanoscale nanoscience nanosystems public health relevance research and development research study response restriction enzyme scale up single molecule stem tool

项目摘要

DESCRIPTION (provided by applicant): 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 Application 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. PUBLIC HEALTH RELEVANCE: Project Narrative The long term goal of our research is to dissect, understand, and control the biology of single cells in complex tissues, such as brain, or in malignant tumors. Furthering this body of work requires that we address an unsolved problem in single cell molecular analysis: the lack of a method to routinely, reliably and inexpensively determine global gene transcriptional activity. The nanotechnologies we are developing 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.

描述（由申请人提供）：项目摘要所有现有的全球基因表达测定技术都需要相对较大的分析物。为了将它们与几个或单个单元中存在的微量mRNA一起使用，需要酶扩增[5]。这个过程耗时，技术困难且昂贵。更重要的是，放大过程本身以禁止真正定量分析的方式偏见样本[6]。这与微阵列和DNA测序的特定局限性相结合，没有直接研究单个细胞中转录的直接方法。为了解决这个问题，我们旨在通过原子力显微镜直接鉴定单个基因转录分子（以cDNA的形式）。该应用程序的具体目的是将我们已开发为集成的功能系统中的硬件，软件和化学的构建块结合在一起。我们提出的改进将通过减少小样本转录分析的时间，成本和技术复杂性来显着影响医学；并通过产生易于解释，更好的信息质量来降低生物信息学负担。公共卫生相关性：项目叙事我们研究的长期目标是剖析，理解和控制复杂组织（例如大脑或恶性肿瘤）中单细胞的生物学。进一步的工作要求我们在单细胞分子分析中解决一个未解决的问题：缺乏常规，可靠和廉价地确定全局基因转录活性的方法。我们正在开发的纳米技术将通过减少小样本转录分析的时间，成本和技术复杂性来重大影响医学；并通过产生易于解释，更好的信息质量来降低生物信息学负担。