High-throughput intracellular import-M/C

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.