Part:BBa_K4960040

pPVC promoter->pvc1-pvc12->pvc13_NTD-2*GGGSG-E01 DARPin-2*GGGSG -pvc13_CTD->pvc14-pvc16

A system for targeted delivery of target proteins.

Usage and Biology

Mutant Pvc13-E01-DARPin fused with the DARPin E01 structural domain can enhance the binding ability of PVCs to cells, ultimately enabling targeted delivery of fat proteins. Pvc13-E01-DARPin mutant-based constructs of PVCs particles can specifically target cells with EGFR receptors[1]. In subsequent cell experiments, we used Hela and HEK-293T with high EGFR expression to ensure the targeting of pvc.

(A)

(A) Schematic diagram of the influence of different Linkers on the structure of tail fibers

Special Design

Use the CKGGRAKDC to target fat cells. The different linker(1*GGGSG or 3*GGGSG) on both sides of the CKGGRAKDC can affect the structure and function of the tail protein. By altering the linker on both sides of the CKGGRAKDC, we explored the most suitable structure of the tail protein for recognition and bond. At the same time, because of the existence of linker, we can use Golden Gate to replace the sequences in the middle of linker more efficiently than enzyme cutting and enzyme linking.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7406
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2407
  Illegal BglII site found at 11264
  Illegal BglII site found at 14099
  Illegal BglII site found at 15464
  Illegal BglII site found at 17422
  Illegal XhoI site found at 2691
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 11792
  Illegal NgoMIV site found at 17953
  Illegal NgoMIV site found at 18092
  Illegal AgeI site found at 6704
  Illegal AgeI site found at 9641
  Illegal AgeI site found at 10327
  Illegal AgeI site found at 10468
  Illegal AgeI site found at 11576
  Illegal AgeI site found at 18391
  Illegal AgeI site found at 20079
  
• 1000
  COMPATIBLE WITH RFC[1000]

Tube:pvc1, pvc5, pvc7
Sheath:pvc2, pvc3, pvc4
Sheath terminator:pvc15
Tail fibres:pvc13_NTD-2*GGGSG-E01 DARPin-2*GGGSG -pvc13_CTD
Baseplate:pvc9, pvc11, pvc12
Spike:pvc8, pvc10
Assembly method:3A Assembly

(B) (B) PVC illustration

References

[1]Kreitz J, Friedrich MJ, Guru A, Lash B, Saito M, Macrae RK, Zhang F. Programmable protein delivery with a bacterial contractile injection system. Nature. 2023 Apr.