High-throughput intracellular import-M/C

我们设计了非常小的聚合物珠（比一毫米小1000倍），它们非常小，以至于哺乳动物细胞可以吸收它们。生物活性分子如酶或DNA可以附着在这些珠子上，这些珠子仍然被递送到细胞中。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”可以改变孤立细胞中的过程，使我们能够在真实的时间内探索细胞的内部工作（一个类比是房子（=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）可以用来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表现），这就是细胞表现为的细胞类型！我们将研究的特定细胞（胚胎干细胞）非常特殊，因为它们在理论上可以用于形成任何所需的组织。控制所形成的组织类型在此时实际上是非常成问题的。我们提出的基于珠粒的传递系统将提供一种新的方法来控制这一过程。珠粒可以用多达100种不同的颜色染色，这些颜色可以用来识别单个珠粒，如果每个珠粒携带不同的生物分子，我们就知道它可以做什么，当我们在显微镜下观察时，它在哪里做。这使得100种不同的RNAi可以连接到100种不同的珠子上，然后可以在一个实验中使用这些珠子来观察100种不同的生物实验（关闭100种不同的基因）。下一步是组合筛选，将两个珠子（不同种类的）放入单个细胞中，从而允许研究100 x100个组合。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，干细胞如何发育。在未来，这将允许开发新的基于干细胞的疗法来治疗或治愈疾病。

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.