Analytical platform for genes and gene regulatory modules

基因和基因调控模块分析平台

• 批准号: 389901-2010
• 负责人: Agellon, Luis
• 金额: $ 3.91万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2009
• 资助国家: 加拿大
• 起止时间: 2009-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=443055
• 关键词: Analytical; platform; genes; gene; regulatory

Changes in "gene activity" (gene expression) that affect the way cells function can be seen as changes in the amount and/or activity of proteins that the genes code for. Cells use some proteins for structure, storage or binding, and some proteins (enzymes) for carrying out biochemical reactions (enzyme activities). The amount of protein/ enzyme activity in the cell reflects the expression status of its gene in the cell. A biochemical test detects changes in the characteristics of the test liquid relative to the amount or activity of proteins/ enzymes in the sample being tested. A microplate reader is used for this purpose. The microplate allows many tests to be done at the same time, and the microplate reader measures the changes in characteristics in the test liquid. Today tests are available for many proteins and enzymes. Older versions of the reader can measure changes in characteristics of the test liquid based on absorption of visible and UV light; modern versions can detect changes in more characteristics (e.g., based on luminescence or fluorescence) in minute amounts of test liquids. Some of the methods used to understand how gene expression is regulated take advantage of the fact that purified DNA (what genes are made of) can be manipulated and reintroduced into cells. A new method has been worked out for efficient introduction of DNA into cells, especially "primary cells" (which are more like the cells in the body, unlike the cancer cells that are widely used by many laboratories today). We are proposing a resource that dramatically improves the efficiency of the way we put genes back into cells AND analyze how the genes respond to specific metabolic states or how they influence the metabolic function of the recipient cells, which helps us to understand better how gene activity is controlled and what genes do in cells. The resource is made up of two instruments: one allows efficient transfer of DNA into cells (especially primary cells), the other allows versatile and sensitive measurements of the activities coded for by genes. This resource will significantly enhance and extend the capabilities of 6 laboratories that have independent and collaborative research programs as well as train individuals who will become tomorrow's professionals.

影响细胞功能的“基因活性”（基因表达）的变化可以被视为基因编码的蛋白质的量和/或活性的变化。细胞利用一些蛋白质进行结构、储存或结合，利用一些蛋白质（酶）进行生化反应（酶活性）。细胞中蛋白质/酶活性的量反映了其基因在细胞中的表达状态。生物化学测试检测测试液体的特性相对于被测试样品中蛋白质/酶的量或活性的变化。酶标仪用于此目的。微孔板允许同时进行多项测试，酶标仪测量测试液体中的特性变化。今天的测试可用于许多蛋白质和酶。旧版本的读取器可以基于可见光和UV光的吸收来测量测试液体的特性的变化;现代版本可以检测更多特性的变化（例如，基于发光或荧光）。一些用来理解基因表达是如何调节的方法利用了这样一个事实，即纯化的DNA（基因的组成）可以被操纵并重新引入细胞。一种新的方法已经被开发出来，用于有效地将DNA引入细胞，特别是“原代细胞”（它更像体内的细胞，而不像今天许多实验室广泛使用的癌细胞）。我们正在提出一种资源，可以大大提高我们将基因放回细胞的效率，并分析基因如何响应特定的代谢状态或它们如何影响受体细胞的代谢功能，这有助于我们更好地了解基因活性是如何控制的，以及基因在细胞中的作用。该资源由两种仪器组成：一种允许将DNA有效转移到细胞（特别是原代细胞）中，另一种允许对基因编码的活动进行多功能和灵敏的测量。该资源将显著增强和扩展6个实验室的能力，这些实验室拥有独立和协作的研究项目，并培养将成为未来专业人士的个人。