Magnet Assisted Transfection

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The Magnet Assisted Transfection (English 'Magnet-assisted transfection', MATra for short , also magnetofection ) is a biochemical transfection method that uses a magnetic field to smuggle nucleic acids into target cells. Here, iron-containing, magnetic nanoparticles are loaded with DNA, which binds them due to ionic interactions. A magnetic field guides the nanoparticles towards and into the target cells. There the DNA is released.

properties

Magnetofection is used to introduce nucleic acids into cells , e.g. B. in the production of transgenic organisms. In this process, DNA , siRNA , dsRNA , shRNA , mRNA or oligonucleotides are bound via their negative charges to magnetic particles that have previously been coated with cationic lipids or polymers . The magnetic particles loaded with nucleic acids are placed on a cell culture in order to subsequently increase the adsorption of the nucleic acids on the cells by applying a magnetic field and consequently their uptake by endocytosis and pinocytosis .

Magnetic nanoparticles, which are used as carriers for nucleic acids, usually consist of iron hydroxides . These are obtained by precipitating iron-containing acid solutions by adding the appropriate bases . The nanoparticles have a size of approx. 100 nm and are usually coated with a biodegradable polymer . The polymer provides the surface of the iron particles with a positive charge, which is important for the ionic interaction with the DNA.

The positively charged magnetic particles bind the negatively charged DNA. The complex formation occurs relatively quickly. The loaded iron particles are then incubated with the target cells on a magnetic plate. The magnetic field causes the iron particles to reach the surface of the cell membrane of the target cells. Those then ingest the particles by endocytosis or pinocytosis . Once in the cell, the DNA is released and the magnetic particles accumulate in endosomes and / or vacuoles and are converted into normal cellular iron metabolism over time. In most cases, the increased iron concentration in the medium does not lead to any cytotoxic effects.

Magnet Assisted Transfection is a time-saving and relatively new technology for introducing DNA into a target cell. The transfection of adherent mammalian cells and primary cell lines is particularly efficient, but suspension cells and cells from other organisms can also be successfully transfected. A great advantage of this method is the careful handling of cells. Other methods are limited by the possible cytotoxic effects of the lipidic transfection reagent ( lipofection ) or by direct force on the cells ( electroporation , 20–50% dead cells). Furthermore, the directed transport of the magnetic particles increases the transfection efficiency in many cases, especially with small amounts of DNA. Methods such as lipofection only provide a statistical chance of the vector and target cell meeting. Magnet Assisted Transfection does not require a serum- free medium for a successful transfection, which offers another advantage.

Currently (as of 2009) over 150 different cell lines and primary cells have been successfully transfected using this method. Further synergistic effects of the transfection efficiency can result from the combination of lipofection and magnet assisted transfection.

This nanotechnology-based transfection method may offer an alternative to the currently used viral and non-viral vectors for in vivo gene transfer and gene therapy in the future .

Alternatives for gene transfer via Magnetofection are transfection with polyethyleneimine , the lipofection , the electroporation , the gene gun that Sonoporation that microinjection and viral vectors .

Individual evidence

  1. ↑ F. Scherer, M. Anton, U. Schillinger, J. Henke, C. Bergemann, A. Krüger, B. Gänsbacher, C. Plank: Magnetofection: enhancing and targeting gene delivery by magnetic force in vitro and in vivo. In: Gene therapy. Volume 9, Number 2, January 2002, pp. 102-109, ISSN  0969-7128 . doi : 10.1038 / sj.gt.3301624 . PMID 11857068 .
  2. ↑ C. Plank, M. Anton, C. Rudolph, J. Rosenecker, F. Krötz: Enhancing and targeting nucleic acid delivery by magnetic force. In: Expert opinion on biological therapy. Volume 3, Number 5, August 2003, pp. 745-758, ISSN  1471-2598 . doi : 10.1517 / 14712598.3.5.745 . PMID 12880375 .
  3. ↑ L. Mair, K. Ford, M. d. Alam, R. Kole, M. Fisher, R. Superfine: Size-uniform 200 nm particles: fabrication and application to magnetofection. In: Journal of biomedical nanotechnology. Volume 5, Number 2, April 2009, pp. 182-191, ISSN  1550-7033 . PMID 20055096 . PMC 2818021 (free full text).
  4. ↑ JI Schwerdt, GF Goya, MP Calatayud, CB Hereñú, PC Reggiani, RG Goya: Magnetic field-assisted gene delivery: achievements and therapeutic potential. In: Current gene therapy. Volume 12, Number 2, April 2012, pp. 116-126, ISSN  1875-5631 . PMID 22348552 .
  5. ↑ Plank, C., Schillinger, U., Scherer, F., Bergemann, C., Remy, JS, Krötz, F., Anton, M., Lausier, J. and Rosenecker, J. (2003) Biol. Chem ., 384, 737-747.
  6. ↑ iba-go.com
  7. ^ Davis, ME (2002) Curr. Opin. Biotechnol. , 13 (2), 128-131
  8. ^ AJ Mellott, ML Forrest, MS Detamore: Physical non-viral gene delivery methods for tissue engineering. In: Annals of biomedical engineering. Volume 41, Number 3, March 2013, pp. 446-468, ISSN  1573-9686 . doi : 10.1007 / s10439-012-0678-1 . PMID 23099792 .