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Efficient Gene Transfection (efficient + gene_transfection)

Distribution by Scientific Domains

Polymers and Materials Science	80%

Selected Abstracts

Brush-Like Amphoteric Poly[isobutylene- alt -(maleic acid)- graft -oligoethyleneamine)]/DNA Complexes for Efficient Gene Transfection

MACROMOLECULAR RAPID COMMUNICATIONS, Issue 13 2010
Majad Khan
Abstract Synthetic gene delivery vectors, especially cationic polymers have attracted enormous attention in recent decades because of their ease of manufacture, targettability, and scaling up. However, certain issues such as high cytotoxicity and low transfection efficiency problems have hampered the advance of nonviral gene delivery. In this study, we designed and synthesized brush-like amphoteric poly[isobutylene- alt -(maleic acid)- graft -oligoethyleneamine] capable of mediating highly efficient gene transfection. The polymers are composed of multiple pendant oligoethyleneimine molecules with alternating carboxylic acid moiety grafted onto poly[isobutylene- alt -(maleic anhydride)]. The polymer formed from pentaethylenehexamine {i.e., poly[isobutylene- alt -(maleic acid)- graft -pentaethylenehexamine)]} was able to condense DNA efficiently into nanoparticles of size around 200,nm with positive zeta potential of about 28,30,mV despite its amphoteric nature. Luciferase expression level and percentage of GFP expressing cells induced by this polymer was higher than those mediated with polyethyleneimine (branched, 25,kDa) by at least one order of magnitude at their optimal N/P ratios on HEK293, HepG2, and 4T1 cells. In vitro cytotoxicity testing revealed that the polymer/DNA complexes were less cytotoxic than those of PEI, and the viability of the cells after being incubated with the polymer/DNA complexes at the optimal N/P ratios was higher than 85%. This polymer can be a promising gene delivery carrier for gene therapy. [source]

Relating Chemical and Biological Diversity Space: A Tunable System for Efficient Gene Transfection

CHEMBIOCHEM, Issue 12 2008
Liisa D. Van Vliet Dr.
Abstract Polyethyleneimine (PEI), a well-established nonviral transfection reagent, was combinatorially modified with varying proportions of methyl, benzyl, and n -dodecyl groups to create a library of 435 derivatized polymers. Screening of this library for transfection, DNA binding, and toxicity allows systematic correlation of the biological properties of our polymers to their derivatizations. Combinations of derivatizations bring about a 100-fold variation in transfection efficiency between library members. The best PEI derivatives exhibit increases in transfection efficiency of more than 80-fold over unmodified PEI (up to 28±7,% of cells transfected) and rival commercial reagents such as Lipofectamine 2000 (21±10,%) and JetPEI (32±5.0,%). In addition, we can identify compounds that are specifically tuned for efficient transfection in CHO-K1 over Ishikawa cells and vice versa, demonstrating that the approach can lead to cell-type selectivity of at least one order of magnitude. This work demonstrates that multivalent derivatization of a polymeric framework can create functional diversity substantially greater than the structural diversity of the derivatization building blocks and suggests an approach to a better understanding of the molecular underpinnings of transfection as well as their exploitation. [source]

Functionalization of Chitosan via Atom Transfer Radical Polymerization for Gene Delivery

ADVANCED FUNCTIONAL MATERIALS, Issue 18 2010
Yuan Ping
Abstract It is of crucial importance to modify chitosan-based polysaccharides in the designing of biomedical materials. In this work, atom transfer radical polymerization (ATRP) was employed to functionalize chitosan in a well-controlled manner. A series of new degradable cationic polymers (termed as PDCS) composed of biocompatible chitosan backbones and poly((2-dimethyl amino)ethyl methacrylate) (P(DMAEMA)) side chains of different length were designed as highly efficient gene vectors via ATRP. These vectors, termed as PDCS, exhibited good ability to condense plasmid DNA (pDNA) into nanoparticles with positive charge at nitrogen/phosphorus (N/P) ratios of 4 or higher. All PDCS vectors could well protect the condensed DNA from enzymatic degradation by DNase I and they displayed high level of transfectivity in both COS7, HEK293 and HepG2 cell lines. Most importantly, in comparison with high-molecular-weight P(DMAEMA) and ,gold-standard' PEI (25 kDa), the PDCS vectors showed considerable buffering capacity in the pH range of 7.4 to 5, and were capable of mediating much more efficient gene transfection at low N/P ratios. At their own optimal N/P ratios for trasnsfection, the PDCS/pDNA complexes showed much lower cytotoxicity. All the PDCS vectors were readily to be degradable in the presence of lysozyme at physiological conditions in vitro. These well-defined PDCS polymers have great potentials as efficient gene vectors in future gene therapy. [source]

Brush-Like Amphoteric Poly[isobutylene- alt -(maleic acid)- graft -oligoethyleneamine)]/DNA Complexes for Efficient Gene Transfection

Synthesis of Guanidinium-Modified Hyaluronic Acid Hydrogel

MACROMOLECULAR RAPID COMMUNICATIONS, Issue 13 2010
Oommen P. Varghese
Abstract In this report, a new guanidinylating reagent is presented, which was developed without any protection/deprotection strategy and was successfully employed for linking to hyaluronan in aqueous solution. The dually functionalised HA biopolymer bearing guanidinium and hydrazide groups was synthesised to form hydrogel in less than a minute when mixed with aldehyde-modified HA. This hydrogel exhibited higher storage modulus with enhanced stability in PBS when compared to the non-guanidine-containing gel. The gel shift assay showed that this biopolymer formed a stable complex with DNA as well as efficient gene transfection to cells that express HA-receptor CD44. The toxicity studies of this polymer with fibroblast cells revealed that the cells were almost 80% viable after 4,d of incubation at high HA concentration (2.5,×,10,3,M). [source]