Short interfering RNA (siRNA) causes sequence specific gene silencing of mRNA, and has been shown to be a very promising therapeutic agent for a wide range of diseases. The delivery of siRNA faces distinct challenges including stability of the formed nucleic acid complexes, protection of siRNA against degradation by nucleases, and efficient intracellular uptake, and release into the cytosol of the cell to silence the targeted gene. To address these issues we have developed a hybrid collagen/ cell penetrating peptide (CHP) carrier, that contains a triple helical collagen domain (POG) that provides stability, and a cell penetrating domain (R<sub>n</sub>) which contains positively charged arginine residues allowing for internalization into cells. Using isothermal titration calorimetry (ITC) we determined that the CHPs form stoichiometric complexes with siRNA with molar ratios of 1:18 and 1:9 depending on the number of arginines present on the CPP domain. These complexes are able to form highly crystalline nanoparticles approximately 280 nm in size with a narrow size distribution of ±7nm. Using multicolor fluorescence confocal microscopy and flow cytometry we determined that the CHPs effectively deliver and release siRNA into 3T3 Swiss mouse fibroblast cells with higher efficiency than Lipofectamine 2000. The data suggests that our strategy for development of the CHP-siRNA complex can provide an avenue for effective delivery of siRNA.

siRNA Delivery with hybrid collagen/cell penetrating peptide carriers

253rd National Meeting (2017)

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