TY - JOUR
T1 - Polyethylenimine-polyethyleneglycol-bis(aminoethylphosphate) nanoparticles mediated efficient DNA and siRNA transfection in mammalian cells
AU - Patnaik, Soma
AU - Tripathi, Sushil Kumar
AU - Goyal, Ritu
AU - Arora, A.
AU - Mitra, Kalyan
AU - Villaverde, A.
AU - Vázquez, E.
AU - Shukla, Y.
AU - Kumar, P.
AU - Gupta, K. C.
PY - 2011/7/7
Y1 - 2011/7/7
N2 - In an attempt to circumvent toxic effects of branched polyethylenimine (bPEI, 25 kDa), it was crosslinked with varying proportions of a novel linker, PEG 600 -bis(aminoethylphosphate) (PaP), which resulted in the formation of nanoparticles (PPaP) in the range of 61-99 nm. These nanoparticles were found to have significantly lower toxicity in vitro than the native PEI. GFP expression in cells mediated by PPaP (8.1%)/DNA complex was found to be ∼1.1-4.8 folds higher compared to GenePORTER 2™, Lipofectamine™, Superfect™ and native PEI in HeLa, HEK293 and CHO cells. FACS analysis on HeLa cells revealed ∼62% transfected cells, whereas, in the case of the GenePORTER 2™ transfection reagent, transfected cells were found to be ∼36%. Intracellular trafficking in HeLa cells showed a significant population of PPaP (8.1%) nanoparticles and their DNA complex in nucleus after 1 h of treatment. Also, efficient delivery of GFP specific siRNA resulted in ∼71% suppression of the target gene. DNase protection assay revealed that ∼78% of complexed DNA was protected by PPaP(8.1%) nanoparticles even after 2 h of treatment. In vivo transgene expression studies in Balb/c mice showed significantly higher expression in the spleen. The results advocate the potential of PPaP nanoparticles as efficient carriers of nucleic acids in vivo. © The Royal Society of Chemistry 2011.
AB - In an attempt to circumvent toxic effects of branched polyethylenimine (bPEI, 25 kDa), it was crosslinked with varying proportions of a novel linker, PEG 600 -bis(aminoethylphosphate) (PaP), which resulted in the formation of nanoparticles (PPaP) in the range of 61-99 nm. These nanoparticles were found to have significantly lower toxicity in vitro than the native PEI. GFP expression in cells mediated by PPaP (8.1%)/DNA complex was found to be ∼1.1-4.8 folds higher compared to GenePORTER 2™, Lipofectamine™, Superfect™ and native PEI in HeLa, HEK293 and CHO cells. FACS analysis on HeLa cells revealed ∼62% transfected cells, whereas, in the case of the GenePORTER 2™ transfection reagent, transfected cells were found to be ∼36%. Intracellular trafficking in HeLa cells showed a significant population of PPaP (8.1%) nanoparticles and their DNA complex in nucleus after 1 h of treatment. Also, efficient delivery of GFP specific siRNA resulted in ∼71% suppression of the target gene. DNase protection assay revealed that ∼78% of complexed DNA was protected by PPaP(8.1%) nanoparticles even after 2 h of treatment. In vivo transgene expression studies in Balb/c mice showed significantly higher expression in the spleen. The results advocate the potential of PPaP nanoparticles as efficient carriers of nucleic acids in vivo. © The Royal Society of Chemistry 2011.
U2 - https://doi.org/10.1039/c1sm05147d
DO - https://doi.org/10.1039/c1sm05147d
M3 - Article
VL - 7
SP - 6103
EP - 6112
JO - Soft Matter
JF - Soft Matter
SN - 1744-683X
ER -