Part:BBa_K4768008

Split T7 RNA polymerase (Nterm) conjugated to Pertuzumab with a transmembrane linker

Part for Expression of the T7 RNA polymerase (Nterm) conjugated to Pertuzumab with a transmembrane linker ZipA

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 987
  Illegal XbaI site found at 40
  Illegal SpeI site found at 1147
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 987
  Illegal SpeI site found at 1147
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 987
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 987
  Illegal XbaI site found at 40
  Illegal SpeI site found at 1147
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 987
  Illegal XbaI site found at 40
  Illegal SpeI site found at 1147
  Illegal AgeI site found at 1200
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 21
  Illegal SapI.rc site found at 1836
  

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Introduction

Figure 1: NT7-ZipA-Pertuzumab structure.

The CALIPSO part BBa_K4768008 is composed of the N-terminal subunit of the T7 RNA polymerase (residues 1 to 180) fused to the anti-HER2 antibody Pertuzumab through an optimized linker sequence containing a transmembrane domain. This gene is under transcriptional control of an SP6 promoter and T7 terminator.

This part, coupled to the part BBa_K4768007 containing the C-terminal subunit of the T7 RNA polymerase, has been designed to develop a split T7 RNAP-based biosensor capable of recognizing HER-2, an epidermal growth factor that is overexpressed in cancer cells. This biosensor will be incorporated into the liposome membrane to detect cancer cells via HER2-targeting antibodies and activate the expression of a gene of interest inside the liposome.

The HER2-induced T7 RNAP complex was designed from two existing constructs: a split T7 RNAP-based biosensor for the detection of rapamycin and a split luciferase conjugated with antibodies capable of recognizing HER2. We decided to merge the relevant functionalities of these two constructs and created a new biosensor that transduces HER2 binding to gene expression activation.

The part BBa_K4768008 was only studied in silico by molecular modeling to optimize the transmembrane linker for adequate flexibility.

Construction

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Molecular Modeling

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Characterisation

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Conclusion and Perspectives

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References

1. article 1 xxxxxxxx
2. article 2 xxxxxxx