Difference between revisions of "Part:BBa K2796028"
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<partinfo>BBa_K2796028 short</partinfo>
 
<partinfo>BBa_K2796028 short</partinfo>
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<li>Considering that <b>TIL cells</b> can't secret enough exosomes under normal condition, we fuse three genes to increase the number of exosomes (Alenquer, 2015), thus increasing the transfer efficiency of exosomes.</li>
 
<li>Considering that <b>TIL cells</b> can't secret enough exosomes under normal condition, we fuse three genes to increase the number of exosomes (Alenquer, 2015), thus increasing the transfer efficiency of exosomes.</li>
 
<li><b>Exosome booster</b> refers to hSDC4-T2A-STEAP3-P2A-NadB.We identified <b>STEAP3</b> (involved in exosomebiogenesis), syndecan-4(<b>SDC4</b>; supports budding of endosomal membranes to form multivesicular bodies), and a fragment of L-aspartate oxidase  (<b>NadB</b>; possibly boosts cellular metabolism by tuning up the citric acid cycle) as potential synthetic exosome production boosters. Combined expression of these genes significantly increased exosome production.(Ryosuke Kojima et al. 2008)</li>
 
<li><b>Exosome booster</b> refers to hSDC4-T2A-STEAP3-P2A-NadB.We identified <b>STEAP3</b> (involved in exosomebiogenesis), syndecan-4(<b>SDC4</b>; supports budding of endosomal membranes to form multivesicular bodies), and a fragment of L-aspartate oxidase  (<b>NadB</b>; possibly boosts cellular metabolism by tuning up the citric acid cycle) as potential synthetic exosome production boosters. Combined expression of these genes significantly increased exosome production.(Ryosuke Kojima et al. 2008)</li>
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[[File:T--LZU-CHINA--design1.png|600px|thumb|center|]]
 
[[File:T--LZU-CHINA--design1.png|600px|thumb|center|]]
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They combine with each other to boost exosomal transfer efficiency：
 
They combine with each other to boost exosomal transfer efficiency：
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[[File:T--LZU-CHINA--design2.png|600px|thumb|center|]]
 
[[File:T--LZU-CHINA--design2.png|600px|thumb|center|]]
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In previous article, it use IRES (internal ribosome entry site,IRES) to connect three genes, we change it into linkers T2A and P2A to ensure proper expression of three genes.
 
In previous article, it use IRES (internal ribosome entry site,IRES) to connect three genes, we change it into linkers T2A and P2A to ensure proper expression of three genes.
 
Then we use use reporter gene copGFP to manifest the expression of exosome booster with the promoter of 5-HRE-PminCMV.
 
Then we use use reporter gene copGFP to manifest the expression of exosome booster with the promoter of 5-HRE-PminCMV.

Revision as of 02:24, 16 October 2018


Exosome booster

  • Considering that TIL cells can't secret enough exosomes under normal condition, we fuse three genes to increase the number of exosomes (Alenquer, 2015), thus increasing the transfer efficiency of exosomes.
  • Exosome booster refers to hSDC4-T2A-STEAP3-P2A-NadB.We identified STEAP3 (involved in exosomebiogenesis), syndecan-4(SDC4; supports budding of endosomal membranes to form multivesicular bodies), and a fragment of L-aspartate oxidase (NadB; possibly boosts cellular metabolism by tuning up the citric acid cycle) as potential synthetic exosome production boosters. Combined expression of these genes significantly increased exosome production.(Ryosuke Kojima et al. 2008)
    • T--LZU-CHINA--design1.png

    They combine with each other to boost exosomal transfer efficiency：

    T--LZU-CHINA--design2.png

    In previous article, it use IRES (internal ribosome entry site,IRES) to connect three genes, we change it into linkers T2A and P2A to ensure proper expression of three genes. Then we use use reporter gene copGFP to manifest the expression of exosome booster with the promoter of 5-HRE-PminCMV. Here is the final device that we designed for this project.

    2018 LZU-CHINA vesicle attacker.png

    It combines with exosome booster and miR attacker under control of induced promoter.More details and description you can obtain in our wiki!


    Sequence and Features


    Assembly Compatibility:
    • 10
      COMPATIBLE WITH RFC[10]
    • 12
      COMPATIBLE WITH RFC[12]
    • 21
      INCOMPATIBLE WITH RFC[21]
      Illegal BglII site found at 192
      Illegal BglII site found at 1777
    • 23
      COMPATIBLE WITH RFC[23]
    • 25
      INCOMPATIBLE WITH RFC[25]
      Illegal AgeI site found at 2657
      Illegal AgeI site found at 3277
    • 1000
      INCOMPATIBLE WITH RFC[1000]
      Illegal SapI site found at 1367