High-throughput intracellular import-M/C

我们设计了非常小的聚合物珠子(大约小于一毫米的1000倍)，这种珠子如此之小，以至于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仍被输送到细胞内的珠子上。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”可以改变隔离细胞中的过程，使我们能够实时探索细胞的内部运作(比喻为一个人在房子里(=细胞))。这些珠子将允许我们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)。从理论上讲，细胞中的任何基因都可以被关闭，结果是细胞表型(例如，细胞在显微镜下的外观)发生变化(S)，也就是细胞的外观类型！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。要控制形成的组织类型，在这个时候其实是很有问题的。我们提出的基于珠子的递送系统将提供一种新的方法来控制这一过程。珠子可以被染上多达100种不同的颜色，可以用来识别单个珠子，如果每个珠子携带不同的生物分子，我们就知道它能做什么，当我们看着显微镜时它在哪里做。这就允许100个不同的RNAi连接到100个不同的珠子上，然后这些RNAi可以用在一个单独的实验中来观察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，将两个不同种类的珠子放入一个细胞中，从而允许研究100x100的组合能够非常迅速地研究如此多的组合将使我们能够了解更多关于如何控制干细胞发育的知识。在未来，这将允许开发基于干细胞的新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.