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: 7948880
• 负责人: Jason C Reed
• 金额: $ 33.88万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 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/7948880
• 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

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，需要进行酶促扩增 [5]。这个过程耗时、技术困难且昂贵。更重要的是，扩增过程本身会使样品产生偏差，从而无法进行真正的定量分析[6]。再加上微阵列和 DNA 测序的特定局限性，导致没有直接的方法来研究单细胞转录。 为了解决这个问题，我们的目标是通过原子力显微镜直接识别单个基因转录分子（以 cDNA 的形式）。该应用程序的具体目标是将我们开发的硬件、软件和化学构建块组合成一个集成的功能系统。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；并将通过产生更容易解释、质量更高的信息来减轻生物信息学的负担。 公共卫生相关性： 项目叙述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。推进这项工作需要我们解决单细胞分子分析中一个未解决的问题：缺乏一种常规、可靠且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，对医学产生重大影响；并将通过产生更容易解释、质量更高的信息来减轻生物信息学的负担。