Electroporation technique

Agrobacterium-mQdi iQd DNA transfer technique and direct gene transfer into protoplasts on the basis of the electroporation technique have frequently been used for the gene transfer methods in plants. However, these methods were hardly adapted to monocotyledonous plants or to varieties in which regeneration system was not established. These deficiencies found in gene transfer techniques will be largely overcome if pollen could be used as a DNA vector. 

Available methods for carrying DNA into an animal cell vary in efficiency and convenience. Some success has been achieved with spontaneous uptake of DNA or electroporation, techniques roughly comparable to the common methods used to transform bacteria. They are inefficient in animal cells, however, transforming only 1 in 100 to 10,000 cells. Microiqjection—the injection of DNA directly into a nucleus, using a very fine needle—has a high success rate for skilled practitioners, but the total number of cells that can be treated is small, because each must be injected individually. 

Physical methods of gene transfer. Genes can often be transferred without the use of a cloning vehicle. This is especially important for certain plant cells, such as those of cereal grains, for which transfer of genes via the Ti plasmid has been difficult.167 If DNA, which may be in a plasmid, is coprecipitated with calcium phosphate, it can often be taken up directly either by animal cells or by plant protoplasts.168 169 Polycations also facilitate DNA uptake cationic liposomes seem to be especially effective.170 In the widely used electroporation technique a short electrical pulse of a few hundred volts / cm is applied to create transient pores in the plasma membrane through which the DNA can enter a cell.111 8,171 175 Chromosomes can be transferred by cell fusion and either... 

Wang, S., M. Kara, and T.R. Krishnan. 1997. Topical delivery of cyclosporin A co-evaporate using electroporation technique. Drug Dev Ind Pharm 23 657. 

Membrane bioreactors can be easily integrated with other systems, for example, with delivery of drugs or genes to individual cells achieved on the nanoscale using electroporation techniques. In one method developed in a recent patent, a flowthrough bioreactor having an inlet and an outlet connected by a flow chamber and a nanoporous membrane positioned in the flow chamber was used [28]. 

Spencer SC (1991), Electroporation technique of DNA transfection, In Murray EJ (Ed.), Methods in Molecular Biology, vol. 7, Humana Press, Clifton, pp. 45-52. 

To address biological processes at the single cell level in a live organism, electroporation techniques have been developed [5]. They involve electroporation by injection of various molecules (e.g., DNA, RNA or morpholinos) present in a micropipette brought in the close vicinity of the targeted cell (see Fig. 16.1). The technique is invasive, and the amount of electroporated material is unknown. Moreover, because the whole tissue is displaced by the inserted micropipette, the success rate in targeting a specific cell in a live embryo is very low. It would be desirable if a noninvasive technique could be devised to control the rapid activation of a known concentration of biomolecules in a specific cell of a live organism. 

Wang, S. Kara, M. Krishnan, T.R. Topical delivery of cyclosporin a coevaporate using electroporation technique. Drug Dev. Ind. Pharm. 1997, 23, 657-663. 

There are different techniques to overcome the cell membrane barrier and introduce exogenous impermeable compounds, such as dyes, DNA, proteins, and amino acids into the ceU. Some of the methods include lipofection, fusion of cationic liposome, electroporation, microinjection, optoporation, electroinjection, and biolistics. Electroporation has the advantage of being a noncontact method for transient permeabilization of cells (Olofsson et al., 2003). In contrast to microinjection techniques for single cells and single nuclei (Capecchi, 1980), the electroporation technique can be applied to biological containers of sub-femtoliter volumes, that are less than a few micrometers in diameter. Also, it can be extremely fast and well-timed (Kinosita et al., 1988 Hibino et al., 1991), which is of importance in studying fast-reaction phenomena (Ryttsen et aL, 2000). 

Rebersek M, Miklavcic D (2010) Concepts of electroporation pulse generation and overview of electric pulse generators for cell and tissue electroporation. In Pakhomov AG, Miklavcic D, Markov MS (eds) Advanced electroporation techniques in biology and medicine. Lecture notes in physics. CRC PressATaylor and Francis Group, London, chapter 16... 

Pakhomov A, Miklavcic D, Markov M (eds) (2010) Advanced electroporation techniques in biology and medicine. CRC Press, Boca Raton, 507 pp... 

Gene transformation has been demonstrated in black spmce using Agrobacterium-msdoaXeA, microprojectile particle bombardment, and electroporation techniques (KUmaszewska et al, 2003 ... 

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