Agriculture Reference
In-Depth Information
This method is generally used to transfer genes into large cells such as oocytes and
the early embryonic stage (Neuhaus et al. 1987 ; Holmberg and B¨ low 1998 ; Baron
et al. 2001 ; Iba 2002 ; Rakoczy-trojanowska 2002 ; Oard 1991 ; Casas et al. 1995 ).
10.3.7 Silicon Carbide-Mediated Transformation (SCMT)
In SCMT method, silicon carbide fibers are added to a suspension, containing both
plant tissues and the plasmid DNA. It is then mixed rigorously so that DNA-coated
fibers could penetrate through the cell wall (Neuhaus et al. 1987 ; Oard 1991 ; Casas
et al. 1995 ; Holmberg and B ¨ low 1998 ; Rakoczy-trojanowska 2002 ; Asad
et al. 2008 ; Sailaja et al. 2008 ). Although SCMT approach is an easy, fast, and
economical method that can be applied to various plants, this methodology has
disadvantages such as low gene transfer efficiency, can damage cells, and nega-
tively influences their regeneration capability, and the precautions to be taken while
dealing with fibers can lead to serious sicknesses (Asad et al. 2008 ; Sailaja
et al. 2008 ). Moreover, the efficiency of SCMT method is affected by various
variables like fiber size, vortexing time, shape of the vessels used, and the thickness
of plant material (Asad et al. 2008 ).
10.3.8 Liposome-Mediated Gene Transfer
Liposomes are colloidal, vesicular structures consisting of one or more lipid
bilayers that surround an equal number of aqueous compartments (Chen
et al. 2011 ; Shirazi et al. 2011 ). Due to the presence of aqueous core and lipophilic
vesicle structure, the plasmid/gene is expected to be encapsulated in an aqueous
environment. Liposomal gene transfer technique has been successfully tried and
adopted in gene delivery as the novel technique (Deshayes et al. 1985 ).
10.3.9 Gene-Loaded Liposomes Provide Numerous
Advantages
Gene loaded Liposomes provides stability to genes, diminish or reduce the deletion
in the DNA, while used along with the physical techniques, control release pattern
of gene delivery, surface modification of liposome (positively charged) potentiate
the penetration within a cell, show a higher degree of reproducibility, applicable to
a wide range of cell types, and free from cellular toxicity. In this technique, the
mechanism of DNA entry through the protoplast seems to be mediated by endocy-
tosis process of liposome by the cells (Shirazi et al. 2011 ), which may involve the
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