Transfection with magnetic beads

Fast +++ Simple ++ Many fold highter transfection rate than conventional system ++ Single as well as Multiple genes with one transfection +++ Easy Transfection of hard to transfect cells like Monocytes +++

Gene delivery, the process of introducing foreign DNA into host cells, is an important step in the genetic modification and is the basis of modern therapies like CAR T cell genetherapies, vaccines and development of recombinant proteins The key point in the success of this genetic transformation has been the development of a safe, stable and efficient gene delivery system. Over the past few decades, many different gene delivery methods have been developed, which can be divided into viral and non-viral types vector systems. Virus-mediated gene delivery utilizes the ability of a virus to inject its DNA into a host cell. Although viral carriers can achieve relatively high levels of transfection efficiency, they have still from many drawbacks, including potential risks of toxicity or immunogenicity, difficulty of production scale-up, and limited capacity to carry DNA beyond a certain size. Therefore, nonviral carriers like Genekam magnetic beads, which have several advantages such as minimal host immune response, stability in storage, and relative ease of production and scale-up, are being developed as alternative methods of gene delivery. Non-viral approaches include agrobacterium-mediated methods, chemical methods such as lipofection, and physical methods such as microinjection, gene guns, hydrostatic pressure, electroporation, continuous infusion, and sonication. Alongside developments in nanotechnology and molecular biology, nanoparticles with positively charged surfaces have shown promise as non-viral carriers for gene delivery. Superparamagnetic nanoparticles like Genekam magnetic beads have many advantages over other

-non-viral gene delivery systems

-enhanced resistance to digestion

-higher DNA carrying capacity

-more powerful penetration

-lower cost

-the ability to drive stable and efficient expression of target genes when exposed to an external magnetic field..

-single as well as multiple genes can be delivered

-simple to perform

-high rate of transfection

Magnetofection with Genekam magnetic beads is a simple, versatile and highly efficient method of gene transfer that uses magnetic force to promote the uptake of gene vectors associated with cationic magnetic nanoparticles into target cells. Effective gene delivery into the mammalian somatic cell nucleus is a key step in achieving a reproductive single cell clone and enabling the breeding of new varieties of animals using the somatic cell nuclear transfer technique. The application of magnetic nanoparticles as gene carriers has progressed rapidly in many fields and number of publications are on increase.

This system can be used for different plasmids, vectors like adeno, retro and orher virus vectors, antisense as well as PCR products. It can be used on different types of cells like monocytes, CHO, HELA, BHK, Vero, endothelial cells, lymphocytes, fibroblasts, chondrocytes, epithelial cells, renal cells, hepatocytes and many more.

Points to be kept in mind for in vitro magnetofection:

The non-adherent cells are to be centrifuged and one can add 30 ul of magnetic neads DNA complex to the pellet and let it mix for 30 minutes. Pipette them in plate or flask, where the magnetic field will be applied. One can also use coated plates so that these cells adhere to the surface. This is based on experience!

The adherent cells are to be grown upto 60-80% confluency at the time of transfection.

The cells between 2000 to 20000 per ml are needed for transfection. Vector is to be prepared in serum free media and add to the cells. The cells should be washed with PBS to make them free from serum and other unnecessary enzymes, which may interfere during the process. The magnetic field will be applied for 30 minutes to 8 hours depending the cell kinds and experience in incubator. After that media can be changed to serum media.

DNA needed per square centimeter space is around 25 to 250 ng. This can help to calculate the total requirement for experiments.