Journal
JOURNAL OF NANOBIOTECHNOLOGY
Volume 17, Issue -, Pages -Publisher
BMC
DOI: 10.1186/s12951-019-0444-8
Keywords
Gene delivery; Non-viral vector; Small interfering RNA; Transfection; RNA interference
Funding
- NIH [R01DE025393]
- NSF [DMR1505214]
Ask authors/readers for more resources
BackgroundWe developed a non-viral vector, a combination of HIV-1 Tat peptide modified with histidine and cysteine (mTat) and polyethylenimine, jetPEI (PEI), displaying the high efficiency of plasmid DNA transfection with little toxicity. Since the highest efficiency of INTERFERin (INT), a cationic amphiphilic lipid-based reagent, for small interfering RNA (siRNA) transfection among six commercial reagents was shown, we hypothesized that combining mTat/PEI with INT would improve transfection efficiency of siRNA delivery. To elucidate the efficacy of the hybrid vector for siRNA silencing, -actin expression was measured after siRNA -actin was transfected with mTat/PEI/INT or other vectors in HSC-3 human oral squamous carcinoma cells.ResultsmTat/PEI/INT/siRNA produced significant improvement in transfection efficiency with little cytotoxicity compared to other vectors and achieved approximate to 100% knockdown of -actin expression compared to non-treated cells. The electric charge of mTat/PEI/INT/siRNA was significantly higher than INT/siRNA. The particle size of mTat/PEI/INT/siRNA was significantly smaller than INT/siRNA. Filipin III and -cyclodextrin, an inhibitor of caveolae-mediated endocytosis, significantly inhibited mTat/PEI/INT/siRNA transfection, while chlorpromazine, an inhibitor of clathrin-mediated endocytosis, did not inhibit mTat/PEI/INT/siRNA transfection. Furthermore, the transfection efficiency of mTat/PEI/INT at 4 degrees C was significantly lower than 37 degrees C.ConclusionsThese findings demonstrated the feasibility of using mTat/PEI/INT as a potentially attractive non-viral vector for siRNA delivery.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available