DNA
ACP7

Part:BBa_K5056002

Designed by: Yik Teng Wong   Group: iGEM24_HK-Joint-School   (2024-09-19)


6X His-tag - de novo design anticancer peptide-7 - linker - cell-penetrating peptide

De novo anticancer peptide is generated by RFdiffusion and ProteinMPNN-AI, acts against the cancer cell by targeting the KRAS mutation. The peptide is designed to bind to the phosphodiesterase-delta (PDE-delta) to hinder the binding of PDF-delta and KRAS protein. This disable KRAS movement and diffusion in the cytoplasm, and discontinue its spatial distribution on the cell membrane.

Two restriction sites (NdeI and SacI) have been added in both ends for digestion and ligation.

6X His tag has been added at the beginning for purification.

One restriction site, NheI, has been added between 6X His tag and the de novo anticancer peptide for future modification.

A middle linker is added between de novo anticancer peptides and cell-penetrating peptide

Polyarginine Cell-Penetrating Peptide: Peptides, especially intracellular functional peptides that can play a particular role inside a cell, have attracted attention as promising materials to control cell fate. However, hydrophilic materials like peptides are difficult for cells to internalize18. Octaarginine (R8) as a Polyarginine Cell-Penetrating Peptide (CPP) is reported to enhance peptide’s ability to cross the cell membrane and deliver cargo19. Hence, ‘RRRRRRRR’ is used as our signal peptide sequence for penetrating cell membrane and enter the cell.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 61
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None