Magnetotransfection is defined as the delivery of nucleic acids or nucleic acid carriers by magnetic particles mediated by magnetic forces. The carriers can be bound to the magnetic particles, which are usually iron oxide nanoparticles, in most cases connected by non-covalent bonds. The magnetic force can overcome the hydrodynamic force , causing the magnetic carriers to aggregate and/or remain in the raked tissue area. In cell culture, magnetic carriers can be deposited into target cells within minutes. This overcomes the diffusion barrier in nucleic acid delivery, and all carriers are in close contact with the target cell, so that the entry of genetic material into the cell can be synchronized. Nucleic acid delivery is greatly accelerated and the infection efficiency of many vectors is increased. Author: T. Friedman et al, Translator: Wei Qin et al. This experiment is from "Gene Transfer".
Operation method
Nucleic Acid Transport with Magnetic Nanoparticles Move Nucleic Acid Transport with Magnetic Nanoparticles Materials reagents Phosphate Buffer Solution (PBS), HEPES Buffer Solution (HBS), 0.9 % NaCl, or for lentiviruses (H airnet al., 2005)]. 9 % NaCl, or serum-free cell culture medium for vector preparation. Selection of transfection reagents Instruments 37°C cell incubator Microcentrifuge Tubes Permanent magnets Rare earth permanent magnets are the most suitable magnets, e.g. neodymiumiron boron magnets. Manufacturers include IBS Magnet (Berlin, Germany), Dexter Magetic Technologies (Germany), and IBS Magnet (Germany). The most practical magnetic sheets are round, 0.5 cm, 1 cm, 1.5 cm, 2 cm or larger in diameter, and 0.5 cm thick. Rectangular sheets measure 2 cm X 1 cm X 0.5 cm. Note: Rare earth permanent magnets produce a permanent magnetic field and must be handled with care. These magnets can be de-magnetized. storage devices and interfere with the proper functioning of electronic or electromechanical devices such as pacemakers. Methods 1 . Preparation of lmg/m l of magnetic nanoparticles with deionized water. Magnetic nanoparticles are supplied by most manufacturers as liquid suspension solutions at various concentrations (mg/ml). Polycation- and polyanion-coated magnetic particles spontaneously aggregate in salt-containing solutions. 2-Calculate the amount of magnetic nanoparticles needed. For non-viral vectors, use 0.2 to 4 ug of outer-coated magnetic particles per microgram of nucleic acid for delivery; 15 ug of polyethylenimine-coated magnetic particles is sufficient to bind I X l O 10 adenovirus particles. The recommended dose of nucleic acid to be added to different culture vessels and the transfection of non-viral vectors are shown in Table 1, and the magnetic transfection of viral vectors is shown in Table 2. Add the desired amount of magnetic pellet to a microcentrifuge tube. For more product details, please visit Aladdin Scientific website.
Division) operating instructions. For polycation- or polyanion-coated magnetic particles, such as CombiMAG, affinity coupling of the vector is not required. Viral vectors can be affinity coupled and the magnetic particles encapsulated with the appropriate streptavidin protein. Alternatively, the vector can be co-incubated with the magnetic particles [cation-coated magnetic particles are preferred, but anion-coated particles can be used for lentiviruses (H airnet al. 2005)].
(Chicago, Illinois), Bisbell Magnetic product Ltd. (Burton-on-T rent, UK), or amfmagnetiC (Mascotte, Australia). Disks designed and optimized for magnetic transfection are commercially available from OZ
Bioscience (Marseille, France; www.ozbioscience.com) or chemicell (Berlin, Germany; WWW.chemicell.com).
See Table 1 for cell culture vessel selection.
For each magnetic particle and vector type, the ratio of magnetic particles to gene vector must be optimized. See www.ozbiosciences.com for specific optimization methods.
