Part:BBa_K5056011:Design
Original C. militaris biomimetic peptide- linker - cell-penetrating peptide
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 61
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 128
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
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 C. militaris biomimetic peptide for future modification.
A middle linker is added between C. militaris biomimetic 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.
Source
ACP: Chantawannakul J, Chatpattanasiri P, Wattayagorn V, et al. Virtual Screening for Biomimetic Anti-Cancer Peptides from Cordyceps militaris Putative Pepsinized Peptidome and Validation on Colon Cancer Cell Line. Molecules (Basel, Switzerland). 2021 Sep;26(19):5767. DOI: 10.3390/molecules26195767. PMID: 34641308; PMCID: PMC8510206.
Middle linker: Part:BBa_K243005
Cell-penetrating peptide: Hao, M., Zhang, L., & Chen, P. (2022). Membrane internalization mechanisms and design strategies of Arginine-Rich Cell-Penetrating peptides. International Journal of Molecular Sciences, 23(16), 9038. https://doi.org/10.3390/ijms23169038
References
Chantawannakul J, Chatpattanasiri P, Wattayagorn V, et al. Virtual Screening for Biomimetic Anti-Cancer Peptides from Cordyceps militaris Putative Pepsinized Peptidome and Validation on Colon Cancer Cell Line. Molecules (Basel, Switzerland). 2021 Sep;26(19):5767. DOI: 10.3390/molecules26195767. PMID: 34641308; PMCID: PMC8510206.
Hao, M., Zhang, L., & Chen, P. (2022). Membrane internalization mechanisms and design strategies of Arginine-Rich Cell-Penetrating peptides. International Journal of Molecular Sciences, 23(16), 9038. https://doi.org/10.3390/ijms23169038