Difference between revisions of "Part:BBa K2208001:Design"

 
 
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<partinfo>BBa_K2208001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2208001 SequenceAndFeatures</partinfo>
  
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In order to obtain exosomes with adipocyte-targeting ability, we modified natural exosomes using a targeting short peptide. Some studies have shown that tPep, homes to adipocytes by strictly binding to prohibitin that are specifically expressed on white fat tissue cells. Lamp2b (lysosomal-associated membrane protein 2b) is a protein found specifically abundant on the surface of exosomes. We connected tPep to Lamp2b using a glycine-linker to construct our fusion protein and promoted its expression by the cmv promoter, hence, Lamp2b can bring the tPep to the surface of exosomes. Then the tPep can guide exosomes to the specific tissues. Through these modifications, exosomes will be conferred on adipocyte-targeting ability.
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[[File:NJU_China_2017_iGEM_Parts_Description_Figure_2.png]]
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[[File:NJU_China_2017_iGEM_Parts_Description_Figure_1.png]]
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Figure 1. tPep-Lamp2b fusion protein modified exosome, with gene specific siRNAs involved inside
  
 
===Design Notes===
 
===Design Notes===
 
In our design, the tPep is inserted between the signal peptide and the N-end of the Lamp2b so that the fusion protein will fold and work normally. And we make sure the tPep will show up at the surface of exosomes.
 
In our design, the tPep is inserted between the signal peptide and the N-end of the Lamp2b so that the fusion protein will fold and work normally. And we make sure the tPep will show up at the surface of exosomes.
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===Source===
 
===Source===
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Artificially designed.
 
Artificially designed.
  
 
===References===
 
===References===

Latest revision as of 14:31, 27 October 2017


A coding sequence of tPep-Lamp2b fusion protein and position it outside the membrane.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1276
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 199

In order to obtain exosomes with adipocyte-targeting ability, we modified natural exosomes using a targeting short peptide. Some studies have shown that tPep, homes to adipocytes by strictly binding to prohibitin that are specifically expressed on white fat tissue cells. Lamp2b (lysosomal-associated membrane protein 2b) is a protein found specifically abundant on the surface of exosomes. We connected tPep to Lamp2b using a glycine-linker to construct our fusion protein and promoted its expression by the cmv promoter, hence, Lamp2b can bring the tPep to the surface of exosomes. Then the tPep can guide exosomes to the specific tissues. Through these modifications, exosomes will be conferred on adipocyte-targeting ability.

NJU China 2017 iGEM Parts Description Figure 2.png NJU China 2017 iGEM Parts Description Figure 1.png

Figure 1. tPep-Lamp2b fusion protein modified exosome, with gene specific siRNAs involved inside

Design Notes

In our design, the tPep is inserted between the signal peptide and the N-end of the Lamp2b so that the fusion protein will fold and work normally. And we make sure the tPep will show up at the surface of exosomes.

Source

Artificially designed.

References