High-throughput intracellular import-M/C

Lead Research Organisation: Aston University
Department Name: Sch of Engineering and Applied Science

Abstract

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.
 
Description We developed new polymer beads that could be used to take protein molecules into cells and once inside the cells drop off the proteins so that they could be active agents within the cell. This method of delivery is important because it allows the imported protein molecules to avoid being taken up by the cell in the usual way (endocytosis) which normally involves the protein being broken down by a cellular structure called an endosome. Delivering the proteins on beads prevented this breakdown from happening - if you like this could be viewed as using the beads as a Trojan horse to "trick" the cell into taking the proteins inside). We also showed we could kill cells in this way. For example by delivering the protein RNase A into cells it was able break down the natural RNA present within the cells and this caused them to die. Such an approach could ultimately be useful in cancer therapy for example.
Exploitation Route by making new bead types we hope to develop proof of concept results into real therapies - this is a long term aspiration of my research group. Other will be similarly enabled by our results which have been published in the open literature and for the most part are not covered by patent application.
Sectors Chemicals,Healthcare,Pharmaceuticals and Medical Biotechnology

 
Title 12-oxo-PDA mimics as 'green' agrochemicals 
Description Some of the developed technology was discussed with Syngenta. For commercial reasons it was decided not to license the patent after initial evaluation. 
IP Reference WO2010149947 
Protection Patent application published
Year Protection Granted 2010
Licensed No
Impact None
 
Title Quantum dots 
Description Some of the developed technology was discussed with Nanoco Technologies - a start up founded by Prof Paul O'Brien (a collaborator on this grant). For commercial reasons it was decided not to license the patent after initial evaluation. 
IP Reference GB0902564.4 
Protection Patent application published
Year Protection Granted 2009
Licensed No
Impact None