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>
 
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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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               src="https://static.igem.org/mediawiki/2017/b/b6/Amazonas_Brazil_Substitui%C3%A7%C3%A3o_Design.png">
  
 
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              <img class="img-responsive" src="https://static.igem.org/mediawiki/2017/2/23/Amazonas_Brazil_DonorDNA.png">
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               src="https://static.igem.org/mediawiki/2017/5/5a/Amazonas_Brazil_Project_8_2.png">
 
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               src="https://static.igem.org/mediawiki/2017/8/8e/Amazonas_Brazil_Project_9_1.png">  
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             </div>
 
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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">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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