Difference between revisions of "Team:Amazonas Brazil/Project"

 
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         <li><a class="Link-pg" href="https://2017.igem.org/Team:Amazonas_Brazil"><img src="https://static.igem.org/mediawiki/2017/d/d4/T--Amazonas_Brazil--LogoBranco.png"  width="104.04";height="90"; class="Menu_image"></a></li>
 
         <li><a class="Link-pg" href="https://2017.igem.org/Team:Amazonas_Brazil"><img src="https://static.igem.org/mediawiki/2017/d/d4/T--Amazonas_Brazil--LogoBranco.png"  width="104.04";height="90"; class="Menu_image"></a></li>
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         <li><a class="bar_text" href="https://2017.igem.org/Team:Amazonas_Brazil/Model">Modelling</a></li>
 
         <li><a class="bar_text" href="https://2017.igem.org/Team:Amazonas_Brazil/Model">Modelling</a></li>
 
         <li><a class="bar_text" href="https://2017.igem.org/Team:Amazonas_Brazil/InterLab">Interlab</a></li>
 
         <li><a class="bar_text" href="https://2017.igem.org/Team:Amazonas_Brazil/InterLab">Interlab</a></li>
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         <li><a class="bar_text" href="https://2017.igem.org/Team:Amazonas_Brazil/Applied_Design">Design</a></li>
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         <li><a class="bar_text" href="https://2017.igem.org/Team:Amazonas_Brazil/Applied_Design">Applied Design</a></li>
 
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                   <a href="https://2017.igem.org/Team:Amazonas_Brazil/HP/Gold_Integrated">
 
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                   <a href="#Specifications">
 
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                   <a href="https://2017.igem.org/Team:Amazonas_Brazil/Model">
 
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                   <a href="https://2017.igem.org/Team:Amazonas_Brazil/Demonstrate#Assembly">
 
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               <h1 style=" margin: 0;padding-top: 0.1em;" id="Specifications"></h1>
 
               <h1 style=" margin: 0;padding-top: 0.1em;" id="Specifications"></h1>
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<img src="https://static.igem.org/mediawiki/2017/thumb/f/f0/Amazonas_Brazil_Project_1.png/648px-Amazonas_Brazil_Project_1.png">
 
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              <h2 style="margin-top: 1.4em;text-align: center;">Specifications</h2>
 
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              <h3 class="SubTitle">Our proposal is to provide a bacterial genome editing approach based on BioBrick parts assembly easy to engineer as A, B, C… CRISPR</h3>
 
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              <p class="p">
 
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                The scientific community had achieved great results using CRISPR-Cas9 mediated genome editing techniques. While these methods are moving forward, some basic methodological aspects related to this approach could be further improved to expand this scientific
 
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                revolution. </p>
 
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              <p class="p">In prokaryotes, the genome editing based on CRISPR/Cas9 machinery require quite laborious days of work, the system is weakly founded on bioengineering principles, demands lots of steps, each one carrying a part of the CRISPR/Cas9 apparatus.
 
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                In addition, off-target events happen frequently, the machinery post-effect in the cell isn’t entirely known yet, among others. To legitimate this information, we integrated our project design with other iGEM's teams who had worked with
 
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                CRISPR/Cas9 in previous years. You can read more about what we <a href="https://2017.igem.org/Team:Amazonas_Brazil/HP/Gold_Integrated" title="" class="Read">found here</a> </p>
 
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              <p class="p">The decoupling of all these shortcomings paved the way to the project's delineation and we elected to focus on a challenge: the lack of standardization, which is a real-world problem - included but not limited to SynBio. Then, to optimize
 
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                the previous methods, we designed a trustworthy, well functioning standardized framework based on BioBrick parts called: the CRISPeasy toolbox.
 
  
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              src="https://static.igem.org/mediawiki/2017/3/3f/Amazonas_Brazil_Project_Events.png">
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              <img alt="Erro" style="width: 80%;height: 80%;margin-left: 10%"
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               <h1 style=" margin: 0;padding-top: 0.1em;" id="Design"></h1>
 
               <h1 style=" margin: 0;padding-top: 0.1em;" id="Design"></h1>
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              <h2 style="margin-top: 1.4em;text-align: center;">Design</h2>
 
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              <h3 class="SubTitle">Check all the parts involved and our approaches with closer details!</h3>
 
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              <p class="p">
 
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                <h3 class="SubTitle">Cas9 (BBa_K2457001)</h3>
 
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                <p class="p">The standardized Cas9 device under control of AraC_pBAD regulatory module. In the single guide RNA presence, it forms a catalytically active ribonucleoprotein (RNP) complex which cleaves specifics target sequences from DNA, leading to
 
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                  double-strand breaks. Through cell repair pathways, it will lay the road to genomic editing in living cells. </p>
 
  
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                <h3 class="SubTitle">Optimized sgRNA (BBa_K2457002)</h3>
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                <p class="p">This biobrick is constitutively expressed by J23100 promoter and codify an optimized sgRNA chimera composed of a protospacer crRNA with a GG motif at the 3’ end of its target-specific sequence, a tracrRNA with extended duplex length and
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                  a mutation at the thymine 4. It was rationally engineered to improve the editing efficiency of CRISPR-Cas9 machinery. When expressed, it guides the Cas9 nuclease activity through Rosalind-Watson-Crick base pairing complementarity with
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                  the target sequence. </p>
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                <h3 class="SubTitle">RecA (Bba_K2457003)</h3>
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            <!--  <p class="p">Adicionar aqui</p> -->
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                <p class="p">RecA is an essential building block part which leads the homologous recombination AND DNA insertion into the genome. After the target sequence cleavage catalyzed by the Cas9-sgRNA complex, the SOS response machinery is activated. As an
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                  output of the repair pathway, RecA builds a DNA-protein filament which investigates for a homologous template on undamaged DNA sequences, assisting in the invasion process and setting-up the Holliday Junction. This generates a crossover
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                  with the strands, leading to the recombination among the undamaged strand and the broken strand. </p>
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                DONOR DNA
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Latest revision as of 03:52, 2 November 2017

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