Journal ArticleDOI
Different behavior of branched and linear polyethylenimine for gene delivery in vitro and in vivo.
Lionel Wightman,Ralf Kircheis,Vanessa Rössler,Sebastian Carotta,Regina Ruzicka,Malgorzata Kursa,Ernst Wagner,Ernst Wagner,Ernst Wagner +8 more
TLDR
Understanding how non‐viral vectors initiate gene expression could lead to the development of new future vectors with enhanced efficacy in non-viral gene therapy.Abstract:
Background
Efficient gene transfer is a major challenge for non-viral gene therapy. Understanding how non-viral vectors initiate gene expression could lead to the development of new future vectors with enhanced efficacy.
Methods
Linear or branched polyethylenimine (PEI)/DNA complexes were generated in varying salt conditions and their transfection efficiencies were compared in vitro and in vivo using reporter genes, luciferase and green fluorescent protein, and rhodamine labeled DNA (pGeneGrip™).
Results
The transfection efficiency of linear PEI22/DNA in vitro was generally greater than that of branched PEI/DNA when complexes were generated in salt containing buffer. However, PEI complexes generated under salt-free conditions generally had low transfection activity in vitro. In contrast, PEI22/DNA salt-free complexes were highly active in vivo. Branched PEI/DNA and salt containing PEI22/DNA complexes were generally 10–100-fold less active than the salt-free PEI22/DNA complexes. Salt-free PEI22/DNA complexes were small, but subsequently grew into aggregates when salt was added. In contrast, PEI25/DNA complexes remained small even after salt was added under the same conditions. Furthermore, PEI22/pGeneGrip™ complexes formed large aggregates associated with the cell membrane, cytoplasm and nucleus, while branched PEI complexes remained as small distinct particles associated with the cell membrane or in the cytoplasm.
Conclusions
Branched and linear PEI/DNA complexes differ in their ability to transfect cells. The greater efficiency of linear PEI might be due to an inherent kinetic instability under salt conditions. Understanding how to employ this kinetic instability of linear PEI could help in designing future vectors with greater flexibility and transfection efficiency in vivo. Copyright © 2001 John Wiley & Sons, Ltd.read more
Citations
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Journal ArticleDOI
The dawning era of polymer therapeutics
TL;DR: The successful clinical application of polymer–protein conjugates, and promising clinical results arising from trials with polymer–anticancer-drug conjugate, bode well for the future design and development of the ever more sophisticated bio-nanotechnologies that are needed to realize the full potential of the post-genomic age.
Journal ArticleDOI
Non-viral vectors for gene-based therapy
Hao Yin,Rosemary Lynn Kanasty,Ahmed A. Eltoukhy,Arturo J. Vegas,J. Robert Dorkin,Daniel G. Anderson +5 more
TL;DR: The biological barriers to gene delivery in vivo are introduced and recent advances in material sciences, nanotechnology and nucleic acid chemistry that have yielded promising non-viral delivery systems are discussed, some of which are currently undergoing testing in clinical trials.
Journal ArticleDOI
Nonviral Vectors for Gene Delivery
TL;DR: Two nonviral gene delivery systems using either biodegradable poly(D,Llactide-co-glycolide) (PLG) nanoparticles or cell penetrating peptide (CPP) complexes have been designed and studied using A549 human lung epithelial cells.
Journal ArticleDOI
Gene therapy progress and prospects: nonviral vectors.
Takuro Niidome,Leaf Huang +1 more
TL;DR: Improvement of delivery methods together with intelligent design of the DNA itself has brought about large degrees of enhancement in the efficiency, specificity and temporal control of nonviral vectors.
Journal ArticleDOI
Nanostructured materials for applications in drug delivery and tissue engineering
TL;DR: The biological functions of encapsulated drugs and cells can be dramatically enhanced by designing biomaterials with controlled organizations at the nanometer scale.
References
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Journal ArticleDOI
A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine
Otmane Boussif,Frank Lezoualc'h,Maria Antonietta Zanta,Mojgan Mergny,Daniel Scherman,Barbara A. Demeneix,Jean-Paul Behr +6 more
TL;DR: Together, these properties make PEI a promising vector for gene therapy and an outstanding core for the design of more sophisticated devices because its efficiency relies on extensive lysosome buffering that protects DNA from nuclease degradation, and consequent lysOSomal swelling and rupture that provide an escape mechanism for the PEI/DNA particles.
Journal ArticleDOI
PEGylated DNA/transferrin–PEI complexes: reduced interaction with blood components, extended circulation in blood and potential for systemic gene delivery
TL;DR: In tumor bearing mice, application of non-PEGylated complexes through the tail vein resulted in reporter gene expression in tail and lung, but severe toxicity was observed in some mice, while PEGylation of the complexes mediated reporter gene transfer to the tumor without significant toxicity.
Journal ArticleDOI
Tracking the intracellular path of poly(ethylenimine)/DNA complexes for gene delivery.
TL;DR: It is found that complexes attach to cell surfaces and migrate into clumps that are endocytosed PEI, whether administered with or without DNA, undergoes nuclear localization in the form of ordered structures.
Journal ArticleDOI
The influence of polymer structure on the interactions of cationic polymers with DNA and morphology of the resulting complexes
TL;DR: At the concentrations studied, these polymers interact electrostatically with DNA forming a unit structure with toroidal morphology; the extent of aggregation of the unit structures in solution depends upon the characteristics of the individual polymer.
Journal ArticleDOI
Activation of the Complement System by Synthetic DNA Complexes: A Potential Barrier for Intravenous Gene Delivery
TL;DR: The data suggests that, by appropriate formulation of DNA complexes, complement activation can be minimized or even avoided, and should facilitate the search for DNA complex formulations appropriate for reproducible intravenous gene delivery.