Transcription Dynamics in Single Cells

单细胞的转录动力学

• 批准号: 8349511
• 负责人: Daniel Larson
• 金额: $ 38.41万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8349511
• 关键词: Bioinformatics; Biological Assay; Biophysics; Breast Cancer Model; Cells; Chromatin; Cities; Collaborations; Collection; Computer Simulation; Computer software; Coupled; Custom; DNA-Directed RNA Polymerase; Data Set; Development; Disease; Doctor of Philosophy; Estrogens; Eukaryota; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Human Cell Line; Image Analysis; In Vitro; Individual; Journals; Laboratories; Lasers; Malignant Neoplasms; Malignant neoplasm of prostate; Mechanics; Meta-Analysis; Microscope; Microscopy; National Cancer Institute; Oncogenes; Paper; Pattern; Photons; Polymerase; Positioning Attribute; Preparation; Programming Languages; Prokaryotic Cells; Publishing; Qualifying; RNA; RNA Polymerase II; Reagent; Recruitment Activity; Regulator Genes; Research; Signal Transduction; Staging; Systems Biology; Techniques; Technology; Time; Tissues; Transcriptional Regulation; Variant; Work; Writing; base; design; experience; fluorescence microscope; malignant breast neoplasm; single molecule

The Systems Biology of Gene Expression Section in the National Cancer Institute was established in February, 2011. During that time, the primary goal has been to build a cellular biophysics laboratory for interrogating gene expression in single cells. A custom fluorescence microscope was designed, and the initial parts were purchased. The microscope consists of a Zeiss AxioObserver inverted microscope stand coupled to multiple Newport single-mode lasers. The microscope stage is a custom stage built in collaboration with Mad City Labs for real-time single molecule tracking. The cameras are EMCCD cameras from Andor. The microscope is optimized for photon collection sensitivity and low drift. The mechanical construction of the microscope is still in the early days. However, much of the control and analysis software was written during this past year using the IDL programming language. Also during the last year, Murali Palangat, Ph. D. was recruited as a lab biologist. Dr. Palangat has extensive experience with transcription in both prokaryotes and eukaryotes and has a superb track record in developing single-molecule biophysical assays for RNA polymerase II. Dr. Palangat has published a number of papers in leading journals demonstrating the connection between polymerase elongation and gene regulation. He developed an in vitro single molecule preparation for mammalian RNA polymerase which has become the enabling technology for labs around the world. He is uniquely qualified for the position in that he is intimately familiar with transcription and gene regulation, appreciative of the nuances of single-molecule studies, and also practiced at operating and managing a biophysical laboratory. During the summer of 2011, Celine Hong was a summer intern in the section. During her time in the lab, she carried out a meta-analysis on two recently published data sets on breast cancer gene regulation. This work leads to direct predictions about the functional significance of co-regulation of genes during estrogen signaling. The testable hypotheses generated from this bioinformatic analysis will form the basis of several experimental efforts in the section in the next fiscal year.

国家癌症研究所基因表达部分的系统生物学是在2011年2月建立的。在此期间，主要目标是建立一个细胞生物物理学实验室，以询问单个细胞中的基因表达。设计了定制的荧光显微镜，并购买了初始零件。显微镜由Zeiss AxioObserver倒置显微镜支架耦合到多个Newport单模激光器。显微镜阶段是与Mad City Labs合作建造的定制阶段，用于实时单分子跟踪。摄像机是来自Andor的EMCCD摄像机。显微镜针对光子收集灵敏度和低漂移进行了优化。显微镜的机械结构仍在早期。但是，在过去的一年中，使用IDL编程语言编写了许多控制和分析软件。同样在过去的一年中，穆拉利·帕朗加特（Murali Palangat）博士被招募为实验室生物学家。 Palangat博士在原核生物和真核生物中具有丰富的转录经验，并且在开发RNA聚合酶II的单分子生物物理测定方面具有出色的往绩。 Palangat博士在领先的期刊上发表了许多论文，证明了聚合酶伸长和基因调节之间的联系。他为哺乳动物RNA聚合酶开发了一种体外单分子制备，这已成为世界各地实验室的能力技术。他具有独特的资格，因为他对转录和基因调节非常熟悉，对单分子研究的细微差别表示赞赏，并且在运营和管理生物物理实验室方面也实践。 在2011年夏季，席琳·洪（Celine Hong）是该部分的暑期实习生。在实验室期间，她对最近发表的有关乳腺癌基因调节的数据集进行了荟萃分析。这项工作导致有关雌激素信号过程中基因共同调节功能意义的直接预测。 该生物信息学分析产生的可检验的假设将构成下一个财政年度本节中几项实验努力的基础。