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      Mammalian cell transfection: the present and the future

      1 , , 1 , 2

      Analytical and Bioanalytical Chemistry

      Springer-Verlag

      Transfection, Nucleic acid, Gene, Single cell

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          Abstract

          Transfection is a powerful analytical tool enabling study of the function of genes and gene products in cells. The transfection methods are broadly classified into three groups; biological, chemical, and physical. These methods have advanced to make it possible to deliver nucleic acids to specific subcellular regions of cells by use of a precisely controlled laser-microcope system. The combination of point-directed transfection and mRNA transfection is a new way of studying the function of genes and gene products. However, each method has its own advantages and disadvantages so the optimum method depends on experimental design and objective.

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          Most cited references 32

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          The promises and pitfalls of RNA-interference-based therapeutics.

          The discovery that gene expression can be controlled by the Watson-Crick base-pairing of small RNAs with messenger RNAs containing complementary sequence - a process known as RNA interference - has markedly advanced our understanding of eukaryotic gene regulation and function. The ability of short RNA sequences to modulate gene expression has provided a powerful tool with which to study gene function and is set to revolutionize the treatment of disease. Remarkably, despite being just one decade from its discovery, the phenomenon is already being used therapeutically in human clinical trials, and biotechnology companies that focus on RNA-interference-based therapeutics are already publicly traded.
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            Magnetofection: enhancing and targeting gene delivery by magnetic force in vitro and in vivo.

            Low efficiencies of nonviral gene vectors, the receptor-dependent host tropism of adenoviral or low titers of retroviral vectors limit their utility in gene therapy. To overcome these deficiencies, we associated gene vectors with superparamagnetic nanoparticles and targeted gene delivery by application of a magnetic field. This potentiated the efficacy of any vector up to several hundred-fold, allowed reduction of the duration of gene delivery to minutes, extended the host tropism of adenoviral vectors to nonpermissive cells and compensated for low retroviral titer. More importantly, the high transduction efficiency observed in vitro was reproduced in vivo with magnetic field-guided local transfection in the gastrointestinal tract and in blood vessels. Magnetofection provides a novel tool for high throughput gene screening in vitro and can help to overcome fundamental limitations to gene therapy in vivo.
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              Gene therapy progress and prospects: magnetic nanoparticle-based gene delivery.

              The recent emphasis on the development of non-viral transfection agents for gene delivery has led to new physics and chemistry-based techniques, which take advantage of charge interactions and energetic processes. One of these techniques which shows much promise for both in vitro and in vivo transfection involves the use of biocompatible magnetic nanoparticles for gene delivery. In these systems, therapeutic or reporter genes are attached to magnetic nanoparticles, which are then focused to the target site/cells via high-field/high-gradient magnets. The technique promotes rapid transfection and, as more recent work indicates, excellent overall transfection levels as well. The advantages and difficulties associated with magnetic nanoparticle-based transfection will be discussed as will the underlying physical principles, recent studies and potential future applications.
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                Author and article information

                Contributors
                +1-215-8980420 , +1-215-5732236 , eberwine@upenn.edu
                Journal
                Anal Bioanal Chem
                Analytical and Bioanalytical Chemistry
                Springer-Verlag (Berlin/Heidelberg )
                1618-2642
                1618-2650
                13 June 2010
                13 June 2010
                August 2010
                : 397
                : 8
                : 3173-3178
                Affiliations
                [1 ]Department of Pharmacology, University of Pennsylvania Medical School—Pharmacology, 36th and Hamilton Walk, Philadelphia, PA 19104 USA
                [2 ]Penn Genome Frontiers Institute, University of Pennsylvania Medical School—Pharmacology, 36th and Hamilton Walk, Philadelphia, PA 19104 USA
                Article
                3821
                10.1007/s00216-010-3821-6
                2911531
                20549496
                © The Author(s) 2010
                Categories
                Trends
                Custom metadata
                © Springer-Verlag 2010

                Analytical chemistry

                single cell, gene, nucleic acid, transfection

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