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DNA Delivery (dna + delivery)

Distribution by Scientific Domains
Life Sciences50%
Polymers and Materials Science37%

Selected Abstracts

Direct DNA delivery into zebrafish embryos employing tissue culture techniques

GENESIS: THE JOURNAL OF GENETICS AND DEVELOPMENT, Issue 1 2001
Raquel Sussman
Abstract Summary: The production of transfected fish embryos requires expertise in injecting the fertilized eggs and/or expensive equipment for electroporation or microprojectiles. This article demonstrates that by exposure to DNA constructs conjugated with transfecting reagents dechorionated Danio rerio embryos are capable of acquiring extracellular DNA and expressing reporter genes. Embryos incubated with pCMVluc complexed with GeneJammer or GenePORTER expressed luciferase 24,48 h after exposure. pCMVGFP DNA mixed with the same agents generated embryos that exhibited differential patterns of expression of green fluorescent protein (GFP). Embryonic development varied depending on the procedure employed and the reporter gene utilized. Expression of the luciferase gene did not interfere with the subsequent development of the embryos. In contrast, the embryos expressing a high level of GFP were affected, probably due to a very active promoter. These results demonstrate the ease of obtaining transfected fish embryos, which facilitate the mass production of new genotypes and extend the procedure to laboratories with limited resources. genesis 31:1,5, 2001. © 2001 Wiley-Liss, Inc. [source]

Precision Polymers: Monodisperse, Monomer-Sequence-Defined Segments to Target Future Demands of Polymers in Medicine

ADVANCED MATERIALS, Issue 32-33 2009
L. Hartmann
Abstract The established technology platforms of solid-phase-supported oligopeptide and oligonucleotide synthesis can be expanded to access fully synthetic macromolecules, preserving both the monodisperse character and the defined monomer sequence. Precision polymers are sequentially assembled from a library of functional building blocks, enabling one to program interaction capabilities or generate functions by sequence-specific positioning of functionalities. Examples are provided, showing that these monodisperse macromolecules can be conjugated to oligonucleotides, oligopeptides, or poly(ethylene glycol)s. The resulting model systems can contribute to the understanding of complex biomedical-related processes. Due to the absence of chemical and molecular-weight distributions in these multifunctional segments, exact correlation of the monomer sequence and (bio)properties is attainable. This is demonstrated by the design of carrier systems that exhibit fine-tuned interactions with plasmid DNA, actively controlling important steps in DNA delivery and transfection, such as polyplex formation, DNA compression, and release of the cargo. [source]

Microparticles for the delivery of DNA vaccines

IMMUNOLOGICAL REVIEWS, Issue 1 2004
Derek T. O'Hagan
Summary:, DNA vaccines have demonstrated a lack of adequate potency in humans, which has necessitated the exploration of various adjunct technologies. Inefficient delivery of DNA vaccines, particularly to antigen-presenting cells, may be contributing to this lack of potency. One effective means of facilitating delivery of DNA vaccines to APCs is through the use of microparticles. In this article, we review the background and rationale for microparticles as a vaccine delivery system, data demonstrating their utility and mode of action for DNA delivery, and the prospects for their development. [source]

Combinatorial Modification of Degradable Polymers Enables Transfection of Human Cells Comparable to Adenovirus,

ADVANCED MATERIALS, Issue 19 2007
J. Green
End-modified poly(,-amino ester)s, easy-to-synthesize degradable polymers, are able to deliver DNA to primary human cells at levels comparable to adenovirus and two orders of magnitude better than the commonly used non-viral vector, polyethylenimine. Small structural changes are found to affect multiple steps of gene delivery including the DNA binding affinity, nanoparticle size, intracellular DNA uptake, and final protein expression. In vivo, these polymer modifications enhance DNA delivery to ovarian tumors. [source]

Poly(glycoamidoamine)s: Cationic glycopolymers for DNA delivery

JOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 24 2006
Theresa M. Reineke
Abstract Polymer science is playing an exciting role in inspiring and advancing novel discoveries in the area of genetic drug delivery. Polymeric materials can be synthesized and chemically tailored to bind and compact nucleic acids into viral-like nanoparticles termed polyplexes that can deliver genetic materials into cells. This article highlights our work in this area to synthesize and study a novel class of cationic glycopolymers that we have termed poly(glycoamidoamine)s (PGAAs). The design of these materials has been inspired by many previous works in the literature. Carbohydrate comonomers have been incorporated into these structures to lower the toxicity of the delivery vehicle, and oligoamine moieties have been added to yield a cationic backbone that facilitates strong DNA binding, compaction, cellular uptake, and delivery of genetic material. PGAAs have been designed to vary in the carbohydrate size, the hydroxyl number and stereochemistry, the amine number, and the presence or absence of heterocyclic groups. Through structure,bioactivity studies, we have discovered that these materials are highly biocompatible, and each specific feature plays a large role in the observed delivery efficacy. Such structure,property studies are important for increasing our understanding of how the polymer chemistry affects the biological activity for the clinical development of polymer-based therapeutics. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 6895,6908, 2006 [source]

Sequential model of phage PRD1 DNA delivery: active involvement of the viral membrane

MOLECULAR MICROBIOLOGY, Issue 5 2002
A. Marika Grahn
Summary DNA translocation across the barriers of recipient cells is not well understood. Viral DNA delivery mechanisms offer an opportunity to obtain useful information in systems in which the process can be arrested to a number of stages. PRD1 is an icosahedral double-stranded (ds)DNA bacterial virus with an internal membrane. It is an atypical dsDNA phage, as any of the vertex spikes can be used for receptor recognition. In this report, we dissect the PRD1 DNA entry into a number of steps: (i) outer membrane (OM) penetration; (ii) peptidoglycan digestion; (iii) cytoplasmic membrane (CM) penetration; and (iv) DNA translocation. We present a model for PRD1 DNA entry proposing that the initial stage of entry is powered by the pressure build-up during DNA packaging. The viral protein P11 is shown to function as the first DNA delivery protein needed to penetrate the OM. We also report a DNA translocation machinery composed of at least three viral integral membrane proteins, P14, P18 and P32. [source]

An alternative method for delivering exogenous material into developing zebrafish embryos

BIOTECHNOLOGY & BIOENGINEERING, Issue 6 2007
Vikram Kohli
Abstract Non-invasive manipulation of multicellular systems is important for medical and biological research. The ability to introduce, remove, or modify molecules in the intracellular environment is pivotal to our understanding of cellular structure and function. Herein, we report on an alternative method for introducing foreign material into developing embryos using the application of femtosecond (fs) laser pulses. When intense fs laser pulses are focused to a sub-micron spot, transient pores are formed, providing a transport pathway for the delivery of exogenous material into embryonic cells. In this study, zebrafish embryos were used as a model system to demonstrate the non-invasiveness of this applied delivery tool. Utilizing optically induced transient pores chorionated and dechorionated zebrafish embryos were successfully loaded with a fluorescent reporter molecule (fluorescein isothiocyanate), Streptavidin-conjugated quantum dots or DNA (Simian-CMV-EGFP). Pore formation was independent of the targeted location, with both blastomere-yolk interface and blastomere pores competent for delivery. Long-term survival of laser manipulated embryos to pec-fin stage was 89% and 100% for dechorionated and chorionated embryos, respectively. To our knowledge, this is the first report of DNA delivery into zebrafish embryos utilizing fs laser pulses. Biotechnol. Bioeng. 2007;98: 1230,1241. © 2007 Wiley Periodicals, Inc. [source]

Bacterial delivery of functional messenger RNA to mammalian cells

CELLULAR MICROBIOLOGY, Issue 5 2005
Christoph Schoen
Summary The limited access to the nuclear compartment may constitute one of the major barriers after bacteria-mediated expression plasmid DNA delivery to eukaryotic cells. Alternatively, a self-destructing Listeria monocytogenes strain was used to release translation-competent mRNA directly into the cytosol of epithelial cells, macrophages and human dendritic cells. Enhanced green fluorescent protein (EGFP)-encoding mRNA, adapted for translation in mammalian cells by linking an IRES element to the 5,-end of the egfp coding sequence, was produced by T7 RNA polymerase in the carrier bacteria upon entry into the cytosol where the mRNA is efficiently released from the lysed bacteria and immediately translated in eukaryotic host cells. Besides the much earlier expression of EGFP being detectable already 4 h after infection, the number of EGFP expressing mammalian cells obtained with this novel RNA delivery technique is comparable to or , especially in phagocytic cells , even higher than that obtained with the expression plasmid DNA delivery strategy. Accordingly, bacteria-mediated delivery of ovalbumin-encoding mRNA to macrophages resulted in efficient antigen processing and presentation in vitro indicating that this approach may also be adapted for the in vivo delivery of antigen-encoding mRNA leading to a more efficient immune response when applied to vaccine development. [source]