PEPFECT
Transfection reagent PepFect
PepFect is a proprietary transfection reagent based on a formulation of chemically modified cell-penetrating peptides specifically developed for condensing oligonucleotides into nanoparticles for intracellular delivery.
PepFect was invented and developed by a consortium of scientists at Stockholm University (Stockholm, Sweden) and some industrial partners.
PepFect is commercialized here by its original inventors.
PepFect - a transfection reagent for oligonucleotide delivery
Unlike many other transfection reagents, PepFect is able to transfect nearly 100% of the cell population and efficiently release the complexed oligonucleotides from endosomes, rendering the delivered oligonucleotides active at their respective targets. Chemical modifications in PepFect ensure that the formulation is stable and active in various cell culture conditions.
PepFect efficiently delivers different types of oligonucleotides such as antisense, siRNA, miRNA and plasmids to different types of cells in culture including HeLa pLuc705, A549, mdx mouse myotubes, a cell culture model of Duchenne’s muscular dystrophy (DMD), and others. Non-covalent PepFect - oligonucleotide nanocomplexes induce splice-correction at rates higher than several other transfection agents on the market and remain active in the presence of serum.
PepFect - oligonucleotide nanocomplexes are taken up by cells.
PepFect nanocomplexes with Cy5-labeled oligonucleotides and 10 kDa A488-Dextran were co-incubated with A549 cells. A major co-localization is seen overall, presented by the appearance of yellow colored vesicles (co-localization of green marked Dextran and red marked PepFect nanocomplexes). The white arrows point out some preeminent co-localization vesicles.
Several Z-slices per image were taken which allowed to confirm the co-localization both in X,Y and Z plane. The image shows one of the Z planes with the highest degree of fluorescence intensity. The fluorescence imaging was overlaid on top of the Differential Interference Contrast image (DIC) in order to have a relative view of the cell together with the labeled vesicles.
Comparison between PepFect and some other transfection agents.
Each reagent was used in accordance with manufacturer's protocol. PepFect was used at molar ratio 1:5. Results are presented as fold increase in luminescence over untreated cells. The values represent the mean of at least three experiments performed in duplicate.
Bars from the left:
1) RNAiMAX; 2)Oligofectamine; 3) jetPEI;
4) Lipofectamine 2000; 5) PepFect.
HeLa pLuc705 cells (5×104) were seeded 24 h prior to experiments into 24-well plates. Cells were transfected for 24 h using 200 nM splice-correcting oligonucleotides in full serum media.
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