High-throughput intracellular import-M/C

高通量细胞内导入-M/C

• 批准号: EP/D038642/1
• 负责人: Paul O'Brien
• 金额: $ 33.8万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2006
• 资助国家: 英国
• 起止时间: 2006 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FD038642%2F1
• 关键词: throughput; intracellular; import

We have designed very small polymer beads (about 1000 times smaller than a millimetre), which are so small that mammalian cells can take them up. Biologically active molecules like enzymes or DNA can be attached onto these beads which are still delivered into the cells. Once in the cells the biological molecules can interact with the cell and enable us to study it. Processes in the isolated cell can be changed by this 'intracellular delivery' which allows us to explore the internal working of the cell in real time (an analogy is a person (= bead) inside a house (= cell)). These beads will allow us to study many different aspects of the way cells work, For example some very special nucleic acids (called RNAi) can be used to shut down specific genes (which control function) within the cells. In theory any gene in a cell could be turned off with the result being change(s) in cell phenotype (e.g. how a cell appears under a microscope), that is the type of cell which the cell appears as! The particular cells we will study (embryonic stem cells) are very special as they can in theory be used to form any desired tissue. To control the tissue type formed is actually very problematic at this time. The bead-based delivery systems we propose will offer a new approach to control this process.The beads can be dyed with up to 100 different colours which can be used to identify an individual bead and if each bead carries a different biological molecule we know what it can do and where it does it when we look down a microscope. This then allows 100 different RNAi's to be attached to the 100 different beads and these can then be used in a single experiment to look at 100 different biological experiments (shutting down 100 different genes). The next step is a combination screen where two beads (of different sorts) are placed into a single cell thus allowing 100x100 combinations to be studied Being able to study so many combinations very rapidly will allow us to learn more about how to control, how stem cells develop. In the future this will allow new stem cell-based therapies to be developed to treat or cure diseases.

我们设计了非常小的聚合物珠（大约比毫米小 1000 倍），它们小到哺乳动物细胞可以吸收它们。酶或 DNA 等生物活性分子可以附着在这些珠子上，并仍被输送到细胞中。一旦进入细胞，生物分子就可以与细胞相互作用，使我们能够研究它。这种“细胞内传递”可以改变分离细胞中的过程，这使我们能够实时探索细胞的内部工作（类比是房子（=细胞）内的人（=珠子））。这些珠子将使我们能够研究细胞工作方式的许多不同方面，例如，一些非常特殊的核酸（称为 RNAi）可用于关闭细胞内的特定基因（控制功能）。理论上，细胞中的任何基因都可以被关闭，结果是细胞表型的变化（例如细胞在显微镜下的外观），即细胞显示的细胞类型！我们将研究的特定细胞（胚胎干细胞）非常特殊，因为理论上它们可以用来形成任何所需的组织。此时要控制所形成的组织类型其实是非常有问题的。我们提出的基于珠子的输送系统将提供一种控制这一过程的新方法。珠子可以染成多达 100 种不同的颜色，可用于识别单个珠子，如果每个珠子携带不同的生物分子，我们知道什么当我们从显微镜下观察时，它能做什么以及它在哪里做什么。这样就可以将 100 种不同的 RNAi 附着到 100 个不同的珠子上，然后可以在单个实验中使用它们来观察 100 个不同的生物实验（关闭 100 个不同的基因）。下一步是组合筛选，将两个珠子（不同种类）放入单个细胞中，从而允许研究 100x100 的组合。能够非常快速地研究如此多的组合将使我们能够更多地了解如何控制、如何干细胞发育。未来，这将允许开发新的基于干细胞的疗法来治疗或治愈疾病。