Part:BBa_K2752011

CPP

This cell-penetrating peptide (CPP) is a cyclic heptapeptide DNP that can be used as a permeation signal for Caco-2 cells. Its amino acid sequence is AC-DNPGNET-CGGGS, in which two cysteines are used to form disulfide bonds.

Sequence and Features

Characterization

HeLa cells were incubated with supernatant which contains GFP-cell penetrating peptide(CPP) for one hour. Fluorescence microscopy was used to detect intracellular GFP by FBW.As shown in Figure 1, it was found that each experimental group had weak green fluorescence, and there was no fluorescence in the blank group and the negative control group.

Figure 1. Observing GFP in HeLa Cells.A: Blank control, no GFP, 10-fold fluorescence microscope; B: Negative control, GFP without penetrating peptide, 10-fold fluorescence microscope; C: TAT experimental group, 10-fold fluorescence microscope; D: R8 experimental group, 10-fold fluorescence microscope; E: DNP experimental group, 10-fold fluorescence microscope; F: Pep-1 experimental group, 10-fold fluorescence microscope.

We use SDS-PAGE to test GFP in HeLa cells.The positive control P1, P2 are cells supernatant after disruption which contains GFP; the negative control is the inclusion of cells incubated with GFP which has no penetrating peptide. The blank control is the inclusion of cells incubated with DMEM medium. Comparing four penetrating peptides, it shows that R8 penetrating peptide and cyclic heptapeptide DNP work best.

Figure 2. Results of SDS-PAGE to test GFP in HeLa cells.B: Blank control O：TAT penetrating peptide S：R8 penetrating peptide P：cyclic heptapeptide DNP H: Pep-1 penetrating peptide M：Marker P1: Positive control 1 P2: Positive control 2 N: negative control

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]