Hello everyone. Welcome to our course outline and highlights. But before we start, let's think a little bit about what we would like you to get out of this course. Let's consider a molecule, and I'm showing here the inhibitor, ibrutinib. If we were a chemist, we can think about this in terms of its connectivity, the type of atoms that it has and maybe the topology. You may even want to think how it could be synthesized. But if we were biochemist, then we actually want to understand how specific amino acid residues interact with this molecule. Indeed, a cysteine residue can covalently bind with this molecule, ibrutinib, by forming a covalent bond to a Michael acceptor that medicinal chemists placed in this molecule deliberately. But if we take a step further back, we can realize that this molecule actually interacts with a specific protein, that's bruton's tyrosine kinase. We can see here using an X-ray structure, of which in fact the molecules that we're showing you were part, that this is a specific binder of this particular protein kinase, and that kinase is important in several blood cancers. If we inhibit it, we actually suppress the disease. In fact, this is generally true of many proteins specifically or particularly oncogenes, many of which are indeed kinases. If we over-express oncogenic kinases in cells like I've done here, we can see that those cells grow on top of each other. We can see that because you can see deep blue spots on the top two plates, that's where the cells have lost their typical ability to suppress their growth when they get confined, and they've started to overgrow the plate. Below, we've added a drug and that inhibits the protein kinase that is driving that growth, and then they suppress and they don't make those spots on top of each other. In fact, this is actually what happens in a patient. If we think back to our a bruton's tyrosine kinase inhibitor, ibrutinib, we can imagine that in a patient, that protein complex will be formed and that will help the patient to recover from the disease. In fact, what we've done is we've drawn a line of logic from chemistry to biochemistry, to cell biology, to medicinal chemistry, and all sorts of how drugs work in a person. It's that perspective we would like you to develop while you're in this course, and we're very, very happy to have you here. How are we going to manage to do that? Well, what we're going to do is we're going to follow a line of logic that will build up your understanding in terms of learning about how to combine different types of science together. We'll start off with an introduction, and that will set you all off in the ideas of chemistry, biology, and physics, and you're actually part of the way through that now. We'll move on to our second module, The Spark of Life. This will be two talks about fluorescence. The first module will deal with the devil is in the minutest detail. We'll learn about how to study cells using immunofluorescence, and the second is a ruler over time and space. We'll start to expand our concepts of fluorescence in terms of how we can measure specific signaling pathways. In The Spark of Life, where we look at how to image cells, we'll learn how to take pictures like this. Here, we're looking at three different biological entities, the blue is showing DNA, the green is showing one particular protein, and the red is showing another. The two proteins have been detected using antibodies. We need to know how those antibodies are working and if they're detecting the protein of interest. From this slide, we can see that different cells, even though they're ostensibly the same cell type, express different proteins differently, and that's an important aspect of imaging that we can learn. We also need to be able to look closer at cells and see what's going on in particular locales like the nucleus. Here, we're looking at ZRANB3 and proliferating cell nuclear antigen. ZRANB3 is in green and PCNA is in red. When we merge those two images together, we can see that the spots that you can see in the red and the green images, merge together because we can see yellow spots appearing where the two proteins are in a similar locale, meaning that they likely act in a similar pathway. We'll also look in live animals. Here we're looking at imaging of fluorescent proteins in a zebra fish. This is actually a stress reporter experiment. Of course, if this fish were particularly stressed, we would see it light up bright green, and we can read out what's happening in the fish just by using these simple fluorescent protein assays. We'll then move to the ruler over time and space. We'll use TIRF microscopy to measure the edge. What's happening on the surface of cells or on the surface of a plate, and we'll go into the lab and see specifically how well-trained TIRF microscopy is being carried out in our labs. We'll also then in the second part of the ruler over time and space, look at specific signaling pathways, and we'll look specifically at TORC2 a central nexus of biological signaling. Then we'll move into design and deliver protein design and modification. This will be in two parts, fusion protein design, and then we'll use those concepts to develop sensors, some of which have been developed here. In the fusion protein design, we'll understand why that's important. Here we're looking at the X-ray crystal structure of a ribosome. This consists of approximately 40 different parts, each of which are denoted by the different colors you can see. If we want to make a fusion protein to any one of those particular structures, those individual proteins shown by different colors, we need to understand how making an extension to that particular individual protein will change the whole, and how it will change how it interacts. It's important that you learn those particular concepts before we go and apply fusion protein senses. Here I'm showing a fusion protein sensor that actually reports in live cells on specific kinase activities. When the kinase is active, you can see that the cells are green. However, if you were to add an inhibitor, which I've done on this slide, you can see that the green disappears and the cells become blue. That way we can read out using fluorescence how active our kinase is. Then we'll move into the last two modules. The more the merrier, where we'll amp up the scope of what we're looking at. We'll start off with a module where we look at making life's work of it, and then we'll move to casting wide your net. In the first module, we'll look at photo-uncaging to trigger specific biological processes regulated by specific lipids in specific locales. Here we're showing how you can photo-uncage a particular lipid globally and that up-regulates calcium. We'll contrast that with how locally uncaged lipids not able to trigger signaling. Finally, in the more the merrier, we'll move into the lab, and we'll have an interesting series of videos from Professor Nicola Winssinger where we talk about protein encoded or nucleic acid libraries, and we'll also talk about cell-based high throughput assays. Please join us, we're very happy to have you here, and we're very interested to see how you like our course. We'd also like you to consider the highlights that we have, which are a diverse array of modern chemical biology techniques that we'll talk about. We'll focus however, on a fundamental understanding because we cannot cover all the concepts of chemical biology in this course, but we can give you the tools to understand all the specific concepts that you will encounter in the predominance of your careers. A large number of quizzes have been given to help tailor your understanding to fit current chemical biology understanding. We've also prepared a lot of content using BioRender, which is a professional software which is used commonly in papers. Finally, we have a large amount of bonus features. We've assembled a large amount of experts from many disciplines to give a rounded perspective. It's important that you hear their different perspectives, because we want you to be able to talk the language of chemistry, and biology, and physics altogether. We've used original and unseen data, some of which you've seen in these slides. We've also created special in the laboratory videos, which you've also seen stills off. We have 30 pages of written original material that will help you to get to the information you need without having to go and look online for it. Thank you very much, we're very pleased to have you. Let's go and move further into the introduction and then later on into the course. Thank you.
