 All right. Thank you so much. So my name is Luis Soengson. I graduated here, PhD, Mechanical Engineering, but I also have been working at the VNC Institute in collaboration with Jim Collins on a variety of topics including another presentation that you will see later on in wearable synthetic biology, but this is one that we wanted to present to you because we believe it's actually pretty cool and is this concept of CRISPR-responsive smart materials. As you know, Rachel already described, you know, for the last maybe, you know, 50 years or so, people have really eagerly experimenting with like biomaterials and realities that we have had a ton of advancement, you know, from composites, metals, polymers, you know, synthetic and functionalized materials that at the end of the day accomplish a certain goal. Reality is that still the idea of bringing new biocompatible materials capable of responding to specific biological triggers is still like something that is a priority. And so our very, I guess, peculiar way of addressing this and we don't want to claim necessarily that, you know, we wanted to solve all the materials problem was can we create programmable, nucleic acid programmable materials that somehow can respond to us and to the environment to do interesting things. And the way we sort of thought it was fun to accomplish this is through this idea of embedding programmable DNA nucleases, which is this, you know, CRISPR enzymes, into materials with interesting chemistry to accomplish such a thing. If you guys, you know, if anyone here is interested on these results, all of everything that I'm going to respond, kind of present here, it has already been published in these publication signs that we recently got out. So the basic principle is that, you know, CRISPR enzymes are highly programmable nucleases. So they have a guide that you can program to really detect nucleic acids in a very specific way. Then if you embed those into materials specifically polymers that have either tethered or that are either tethered or cross-linked with DNA, then suddenly you can actuate those materials and change their properties. Specifically, the things that I'm going to show today is a couple of demonstrations that we did across different polymer chemistries, specifically PEG hydrogels, polycretamide DNA gels, and carbon black DNA gels. So just for you to sort of, just for you to kind of like understand a little bit better this diagram, in the first blog that says PEG DNA gels, for example there you can have a matrix that is made of primarily PEG, but you can have the single stranded DNA that has cargos tethered into them. Those, as you will see later, can be, you know, anything from enzymes to fluorophores to small molecules, then then we can cleave in order to make materials that release things into the environment. But in other chemistries such as the middle one, that says polycretamide DNA gels, we can actually have DNA as the fundamental cross-linking unit, so that you can actually trap things inside the material or have a very densely cross-linked material, but the moment that you cleave those through the activity of CRISPR enzymes, which again are programmable, then you can suddenly actuate those materials to swell more or to be more permeable, basically affecting their mechanical properties, at least in the sense of permeability. And the third one that I will discuss today is this idea of other more interesting potentially conductive materials, so these carbon black DNA gels. Carbon black is a material that can be made to be highly conductive, and if you, because of functional groups that it presents, it actually has this capacity to cross-link tightly with single stranded DNA. So just by combining carbon black with DNA, you can actually make these polymers, complex polymers that are conductive, but yet they are mechanically attached together by these DNA strands, hopefully to create potentially electronics that can be modulated by the activity of CRISPR. Alright, so schematically this is what we accomplished, and I'll show you data in a moment. We were able to demonstrate that for these materials, specifically these polymeric hydrogels, we are able to on-demand release cargos, which include enzymes, small molecules, but also cells. We're able to create conductive materials that, upon sensing of nucleic acids in the environment, can detach from electrodes, basically, you know, short circuiting, or in this case open circuiting, you know, electronic circuits to be measured, but also as medium to change permeability, as we discussed previously, in order to do other interesting things, like short circuiting RFID antennas and other things that are kind of more in the realm of devices, to make it more visible of how, like, you could use this for diagnostics and other purposes. So I know that this is kind of like a data-dense slide, but just bear with me in a minute. So here we have this PEC matrix, and again, we're able to tether molecules here. In panel B, for example, you see a fluorophore that is on-demand released from this hydrogel upon the addition of a single stranded DNA that is our target. So in this case, a gel has this molecule, and it has embedded cast-12A enzyme, which is a type of CRISPR enzyme, and it has been programmed to detect a specific target, DNA target. And as you can see, depending on the presence of a specific target, as compared to a scramble DNA in the environment, you can have a very different release profile. We did the same for enzymes, so you can not only imagine to release small molecules into the environment, but also enzymes into the surrounding medium, and that, as other presentations have alluded to, could have very interesting implications, because you can release peroxidases or other things into the surrounding buffer to have more interactions and do more catalytic activity, for example. The interesting part about CRISPR is that it's not only highly programmable, but it's also very orthogonal. So here, for example, we created this hydrogel in panel D to detect four different types of, actually five different genes that are involved in the resistance of methicillin-resistant staphylococcus aureus, which is the super bug that you find in hospitals. And you can see that depending on the different target that you expose this to, you have a different amount of activity, which is really nice. In terms of the electronic circuits, what we did is we created this carbon black DNA hydrogels that we basically just deposited into inter-digitated electrodes. As you can see here, those are just, they look like little dots, dark dots into, you know, in these electrodes. What is interesting is that when you embed these electrodes into an environment that has, for example, the target that you want to detect, these dots just dissolve or detach from the electrodes much more frequently when you have an environment that has that target. So here, for example, you can envision a circuit that is just like stands there, but the moment you interact with it or the environment interacts with that electrode and that environment has a nucleic acid target that you're interested in detecting, then itself the circuit, itself the conductive pathways on those circuits will change. And that's what we have demonstrated here. Obviously, this is not perfect technology. For example, many of these interactions happen at the interface between the inter-digitated electrode and the bulk material, really destroying a bead of hydrogel on itself. It can be a lengthy process for enzymes, but in this case, we're able to achieve these results in a couple of hours, which is interesting. Now, a big application that we thought was interesting was for not only for drug delivery, but also for release of viable cells that, for example, can be immune cells. In this experiment, for example, we created these hydrogels that were able to encapsulate viable mononuclear stem cells on them. And so those, you know, in our publication we have other sort of gels that you can see, but these look very well in the projector. We have these, let's say hydrogels. Those could be potentially implanted into an animal and to a human, and those will actually hold stem cells on them that may be programmed or not to do whatever. But at the end of the day, the idea is that you don't want those cells to be necessarily circulating all the time or at any point, but rather only when certain cues are sort of presented to them. And in this case, nucleic acids were the things that we wanted to detect. So if this is immune cells, you can imagine that you can have, you know, if you have a nucleic acid that is relevant in the detection of certain virus or certain bacteria that you want those immune cells to be released on demand. Also for cancer therapeutics, this seemed to be interesting. And so what we've demonstrated here is that we, you know, if we just present these gels with scramble DNA, you know, don't really dissolve that much in, for example, in this first hour, which is the first section in the left. But as you are presenting those with more and more of this specific DNA trigger, then those gels just dissolve out and those release those cells. And the last section of that panel just shows that kind of like a viability test on the cells just to ensure that, you know, the actual process of releasing them is not killing those cells, which is those green dots that you see there. So, you know, I know that this is all like different layers of complexity, but something that we thought was interesting is, you know, can we, since we have the capacity to create and to sort of dissolve these gels to, can we use these gels in order to do more interesting stuff in terms of flow. And so, you know, there are some people in the last 20 years that have created these stop flow essays where, you know, changing in, you have a paper, a lateral flow in one essay, for example, you have a paper strip where you have flow, but you can stop that flow by making something crosslink along the way during that flow. And that's exactly what we did here. We did this, you know, in literature they are called origami and micro-pads, but basically they are just layers of paper that have been printed with wax to have like almost like channels into them defined. And we can put in different layers of paper, and all these things are paper, you can put different substrates for the crosslinking of these hydrogels. So you can have the precursors, the polymers, you know, you can have dyes, you can have buffer, everything that you need. But what we did here is create this assembly that has all the precursors and everything to create those gels as flow is happening. But where in the presence of a target, the cast enzymes will be activated, basically leaving everything that makes possible for these systems to crosslink, basically allowing for flow to happen if there is a presence of a target and flow to be stopped in the absence of a target. As you can see in the panel on the right, for example, in the presence of the enzyme, you can see in the bottom that buffer flows, and you can see kind of like in this lateral flow section in the bottom that there is this color dye that is flowing through. And you can see the matrix there of the cellulose just, you know, clean in a way, whereas in the upper section, when there is no target and therefore the cast is not activated, then you get hydrogel forming stopping the flow. And so as you can see, you can have just a very, you know, easy colorimetric signal out of these same sort of materials being, you know, broken down or not. But you can also imagine, sorry, measuring, you know, conductivity out of this, because as a buffer is flowing and these buffers are usually, you know, PBS or just, they have salts on them, you can actually think about measuring the conductivity of these micropads as a measurement of like distance of how much buffer flows. That's exactly what we did in order to just almost like solidify this concept that we could use this very intercommunicated like systems, you know, the chemical, the enzymatic asper of CRISPR, the genetics out of it, but also sort of the permeability aspect of our materials and then potential electronic uses. We did a couple of demonstrations where we actually just in colorimetric experiments, which is this panel A, were able to show that we were able to create a very cheap diagnostic for Ebola that was able to detect 11 atom-molar, like up to 11 atom-molar concentrations of Ebola trigger. And that's pretty incredible because, you know, that's pretty much both parking day in where like PCR is. And this is just in a piece of paper, right? And you're seeing kind of like smart materials. So it's an interesting application, this pathway that we're pursuing. But, you know, sometimes people want to have this integrated into electronics to do kind of like more automated reporting. And so I need to stop now. But the idea here is that we integrated this piece of paper also with RFID tags to short circuit a small RFID tag. And that suddenly that's what it hit us because, you know, when you are able to modulate really like materials in this way that are potentially conductive or not, then you can really have these new interfaces between kind of like biology and electronics in a way that we didn't expect to. So again, if anyone is interested about this, everything is sort of published already in science. And we're about to just to release these nature protocols so that anyone can kind of replicate these type of results. I want to thank everyone that was involved in this. And thank you so much.