title: 1,4-Butanediol diglycidyl ether (BDE)-crosslinked PEI-g-imidazole nanoparticles as nucleic acid-carriers in vitro and in vivo.
abstract: Previously reported 1,4-butanediol diglycidyl ether (BDE) crosslinked PEI (branched polyethylenimine, 25 k) nanoparticles (A. Swami, R. Kurupati, A. Pathak, Y. Singh, P. Kumar and K. C. Gupta, A unique and highly efficient non-viral DNA/siRNA delivery system based on PEI-bisepoxide nanoparticles, Biochem. Biophys. Res. Commun., 2007, 362, 835-841) (PN NPs) were reacted with varying proportions of a novel linker, 2-(N-1-tritylimidazol-4-yl)-N-(6-glycidyloxyhexyl)-acetamide (IGA linker, 3), to yield PN-g-imidazolyl nanoparticles (PNIm) with improved transfection efficiency. Here, the IGA linker (3) reacted through an epoxy ring to partially convert the residual 1° and 2° amines present in PN NPs to 2° and 3°, respectively, without altering the total number of amines and additionally incorporating the delocalized positive charge of the imidazolyl moiety. The resulting particles were characterized for their size, zeta potential and DNA complexing ability. PNIm/DNA nanoplexes, in the size range of 120-400 nm, were evaluated for transfection efficiency in HeLa, HEK293 and CHO cell lines, which was found to be ∼11, ∼2-3 and ∼2-17 folds higher than PEI, PN-2 (the best working sample of the PN series) (A. Swami, R. Kurupati, A. Pathak, Y. Singh, P. Kumar and K. C. Gupta, A unique and highly efficient non-viral DNA/siRNA delivery system based on PEI-bisepoxide nanoparticles, Biochem. Biophys. Res. Commun., 2007, 362, 835-841) and commercial transfection reagents tested in this study, respectively. Also, flow cytometric analysis showed ∼78% (ca.∼43% in PN-2) cells transfected with the PNIm 10(6)/DNA complex (the best working sample of the PNIm series) in HEK293 cells. Transfection of GFP specific siRNA in HEK293 cells suppressed the gene expression by ∼90% (ca.∼70% in PN-2). All the cell lines treated with PNIm/DNA nanoplexes showed >90% viability. In vivo gene expression of luciferase enzyme in Balb/c mice showed highest expression in spleen after seven days.
authors.affiliation: Institute of Genomics and Integrative Biology (CSIR), Mall Road, Delhi University Campus, Delhi-110 007, India.
authors.forename: Ritu
authors.initials: R
authors.lastname: Goyal
authors.forename: Ruby
authors.initials: R
authors.lastname: Bansal
authors.forename: Shilpa
authors.initials: S
authors.lastname: Tyagi
authors.forename: Yogeshwer
authors.initials: Y
authors.lastname: Shukla
authors.forename: Pradeep
authors.initials: P
authors.lastname: Kumar
authors.forename: Kailash Chand
authors.initials: KC
authors.lastname: Gupta
mesh_terms: D000818:Animals; D002072:Butylene Glycols / Q000737:chemistry*; D016466:CHO Cells; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004247:DNA / Q000008:administration & dosage*; D003850:Deoxyribonuclease I / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry*; D024202:Electrophoretic Mobility Shift Assay; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D007093:Imidazoles / Q000737:chemistry*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009206:Myocardium / Q000378:metabolism; D053758:Nanoparticles; D009928:Organ Specificity; D010316:Particle Size; D011094:Polyethyleneimine / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000008:administration & dosage*; D013154:Spleen / Q000378:metabolism; D014162:Transfection / Q000379:methods
chemical_list: D002072:Butylene Glycols; D003432:Cross-Linking Reagents; D004337:Drug Carriers; D007093:Imidazoles; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D004247:DNA; C014376:1,4-bis(2,3-epoxypropoxy)butane; D008156:Luciferases; D003850:Deoxyribonuclease I