Hi, we are trying to transfect breast cancer cell lines with several cell penetrating peptide-siRNA complexes but it seems that our peptides are not taken up by the cells.
- We use several CPPs: 8R, 10R, 8R-RGD, 10R-RGD, cRGD-8R, cRGD-10R and complex them to our siRNA for 30min at 37C.
- Based on gel retardation assays, we then selected the optimal molar ratio for transfection.
- When transfecting the 10R-RGD/siRNA complexes in MDA-MB-468 cells, we can observe large circular fluorescent (siRNA) structures floating around but after few hours these disappear and no fluorescent signal can be observed in the cells.
So, basically the 10R-RGD/siRNA complexes are not taken up by the cells.
I would appreciate any advice on how to improve cellular uptake as we are completely stuck with this project at the moment.
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