Difference between revisions of "Part:BBa K2876006"
 
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This part fused the  alpha subunit of a RNAPolIII,BBa_K2876002, (https://parts.igem.org/Part:BBa_K2876002) with a tri-alanine linker for fusion to other proteins. ]
 
This part fused the  alpha subunit of a RNAPolIII,BBa_K2876002, (https://parts.igem.org/Part:BBa_K2876002) with a tri-alanine linker for fusion to other proteins. ]
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Inspired by the yeast two-hybrid system designed iGEM Tsinghua 2017 [3], we fused antibodies and dCas9 proteins to the subunits of the P2H system, creating a versatile platform capable of detecting specific DNA sequences, small molecules, and proteins. Our DNA detection system works with dCas9-protein fusions, which can be targeted to specific DNA sequences (FIGURE 2: DNA detection figure), and is called casP2H (cas9 prokaryotic two hybrid system). Our protein and small molecule system works by fusing single-chain-antibodies specific to different epitopes of a target protein or small molecule, and is called SCAP2H (single chain antibody prokaryotic two hybrid system).
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This detection system is unique because it utilizes transcription. Detection-induced transcription allows genes to be activated by the presence of a user-selected molecule, therefore creating a genetic output to a chosen input. For example, a therapeutic protein could be transcribed in response to a disease biomarker.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 02:40, 18 October 2018


alpha+linker

This part fused the alpha subunit of a RNAPolIII,BBa_K2876002, (https://parts.igem.org/Part:BBa_K2876002) with a tri-alanine linker for fusion to other proteins. ]

Inspired by the yeast two-hybrid system designed iGEM Tsinghua 2017 [3], we fused antibodies and dCas9 proteins to the subunits of the P2H system, creating a versatile platform capable of detecting specific DNA sequences, small molecules, and proteins. Our DNA detection system works with dCas9-protein fusions, which can be targeted to specific DNA sequences (FIGURE 2: DNA detection figure), and is called casP2H (cas9 prokaryotic two hybrid system). Our protein and small molecule system works by fusing single-chain-antibodies specific to different epitopes of a target protein or small molecule, and is called SCAP2H (single chain antibody prokaryotic two hybrid system).

This detection system is unique because it utilizes transcription. Detection-induced transcription allows genes to be activated by the presence of a user-selected molecule, therefore creating a genetic output to a chosen input. For example, a therapeutic protein could be transcribed in response to a disease biomarker.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 8
    Illegal NotI site found at 774
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 480
  • 1000
    COMPATIBLE WITH RFC[1000]