Difference between revisions of "Team:USP UNIFESP-Brazil/Project"

 

<p>MEMBERS</p>

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<div class="small-3 columns membro" data-nome="PROJECT" data-img="https://static.igem.org/mediawiki/2016/d/d3/T--USP_UNIFESP-Brazil--Allan.jpg">

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<p class="black">DESCRIPTION</p>

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<div class="small-3 columns membro" data-nome="EXPERIMENTS" data-bio=" data-img="https://static.igem.org/mediawiki/2016/9/99/T--USP_UNIFESP-Brazil--AndreMaizel.jpg">

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<p class="black">EXPERIMENTS</p>

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<div class="small-3 columns membro" data-nome="PROOF OF CONCEPT" data-img="https://static.igem.org/mediawiki/2016/0/0d/T--USP_UNIFESP-Brazil--Brayan.jpg">

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<p class="black">PROOF OF CONCEPT</p>

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<div class="small-3 columns membro" data-nome="RESULTS" data-img="https://static.igem.org/mediawiki/2016/7/7d/T--USP_UNIFESP-Brazil--BrunoRA.jpg">

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<p class="black">RESULTS</p>

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<div class="small-3 columns membro" data-nome="NOTEBOOK" data-img="https://static.igem.org/mediawiki/2016/2/26/T--USP_UNIFESP-Brazil--Caua.jpg">

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<p class="black">NOTEBOOK</p>

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<div class="small-3 columns membro" data-nome="SAFETY" data-img="https://static.igem.org/mediawiki/2016/0/06/T--USP_UNIFESP-Brazil--Clarissa.jpg">

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<p class="black">SAFETY</p>

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<div class="small-3 columns membro" data-nome="MODELLING" data-img="https://static.igem.org/mediawiki/2016/7/7d/T--USP_UNIFESP-Brazil--Danilo.jpg">

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<p class="black">MODELLING</p>

 

Some studies show silk can be compared to a synthetic polymer called Kevlar, that is made of aramid and is used to make body armor (Lewis). Silks are polymers composed of proteins. These proteins and their genes present specific repetitive sequences that are associated with silk properties, making it possible to handle it in order to obtain the most suitable material for each application. Moreover, some studies suggest that the terminal domains of silk proteins play important roles in the polymerization process. In this way, it can be possible to add N- and C- terminal domains in several interesting proteins or motifs and spun it with native proteins to obtain a customized silk. </p>

                                 <p class="black">The size of genes, their repetitive sequences and high content of Cytosine and Guanine establish a challenge to many expression systems. However Chlamydomonas reinhardtii has naturally high CG content in its genome and it could be able to deal with this problem, it can also perform post translational modifications and it can secrete complex proteins, easing the purification process. Furthermore, it exhibits low cost production, rapid growth, scalability and stable transgenic line generation, all desired characteristics to an expression system (Wijffels, Rosenberg). It is important to highlight the low cost of microalgae cultivation, while production of antibodies in mammalian cells presents an average cost about US$150.00/per gram of raw materials and plants US$0.05/per gram (Dove), Maylfield has estimated that microalgae can reach US$ 0.002/liter. Chlamydomonas reinhardtii also is Generally Recognized As Safe (GRAS), that is, it has low risk of being contaminated by virus, prions and bacterial endotoxins (Mayfield). Easy purification and low risk of contamination are very significant issues when we are talking about making products to biomedical applications.</p>

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                                 <p class="black">From this idea we thought: “Silk is a very interesting material and could be expressed in Chlamydomonas reinhardtii, a model organism that has many advantages and that remains underexplored. What could be put in place of a GFP to express in microalgae? What biomolecule could take advantage of this amazing silk feature? It has to work while immobilized... Why not enzymes? There are many studies about enzymes immobilization, enzymes are very appropriate and useful in many applications around the world!”. But we knew it could not to be any enzyme linked to silk, we knew it need to address technical, social and economic issues. So we read that spider silk has been studied to many biomedical applications, like the treatment of burn victims. Then we studied about burns in order to find an interesting enzyme that could solve some problem and we learned that one of the most important problems affecting burn victims is sepsis. In this way, we sought for antimicrobial enzymes and we found the endolysins. Endolysins are very interesting enzymes because they are able to combat multiresistant bacterias and they are specific, avoiding problems like the antibiotic resistance. </p>  

<p class="black">In our project, we propose to explore the modular characteristic of spider silk proteins, by using it as an immobilization support to other proteins. We were inspired by UCLA’s iGEM Team’s project in 2014 and 2015, where they presented the idea of using silk fibers to integrate other functional proteins to the silk’s structure. We tried to expand on this concept by expressing proteins with antimicrobial activity, enzybiotics (Fig.1). By combining these proteins and their properties, we tried to tackle a major problem with wound dressings for burn victims.</p>

<p class="fig-label">Figure 2: Project overview. Schematic representation of spider web structure from macro to nano scale. A representation of: enzybiotic protein from a bacteriophage; a spider silk protein with repetitive domains and N and C terminals; host expression system Chlamydomonas reinhardtii and a chimeric protein envisioned in this project; and the final product, a biopatch produced from recombinant silk proteins and chimeric proteins.</p>

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<p class="black">Immobilization techniques are applied to a wide range of treatments and processes, from medical applications to biotransformations in industrial plants. This stabilization is normally achieved by protein binding to a scaffold (Liese and Hilterhaus 2013). Recent studies explored spider silks as a possible immobilization support (Blüm et al. 2013, Monier 2013).  Spider silk is known mainly for its tensile strength and fracture resistance, but also exhibits elasticity, adhesion, biocompatibility and low degradation. Its strength can be compared to Kevlar synthetic polymer, which is composed of aramid and is used in for manufacturing body armor (Lewis 2006). Furthermore, medical applications are possible due to its biocompatibility and biodegradability, as coating for implants and transplanted organs, drug delivery and scaffolding for cell lines (Lewis 2006, Hardy et al. 2008, Kluge et al. 2008).</p>

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