Transfection matrices

Fig. 1.11 Applications of LDHs as (A) non-viral vector in gene therapy for transfection of DNA to the cell nucleus, and (B) as matrix for enzymes immobilization in the development of biosensors.

It should be emphasized at this point that the use of physicochemical methods is so far the only way to demonstrate the import of transgene DNA into the mitochondrial matrix in living mammalian cells. The unavailability of a mitochondria-specific reporter plasmid designed for mitochondrial expression severely hampers current efforts toward the development of effective mitochondrial expression vectors. Although any new nonviral transfection system (i.e., cationic lipids, polymers, and others) aimed at the nuclear-cytosolic expression of proteins can be systematically tested and subsequently improved by utilizing anyone of many commercially available reporter gene systems, such a methodical approach to develop mitochondrial transfection systems is currently impossible. 

Immunization with the monoclonal AB reactive against the 18 kDa major surface protein in the oncosphere of T. saginata, HP6, confers protection in cattle (Harrison and Rarkhouse, 1986). The HP6-encoding gene of T. saginata has been cloned and transfected into normal rat kidney cells. Localization studies revealed HP6 in the endoplasmic reticulum, the Golgi apparatus and at the cell surface (Bonay et al., 2002). The deduced amino acid sequence of HP6 was similar to the mammalian extracellular matrix protein,... 

Consequently, San Juan and coworkers chemically cross-linked DEAE-pullulan and synthesized a cationic, 3D pullulan matrix that could be loaded with pDNA (pSEAP) and function as a delivery vehicle (Fig. 6) [82], In vitro transfection and... 

An HA-rich matrix often correlates with a worse clinical prognosis. Metastatic potency is often enhanced in animal model systems when tumor cells are transfected with HAS genes. Inhibition of HAS gene expression with anti-HAS constructs reduces tumor adhesion and malignant potential [123,124]. 

Figure 3 Schematic representation of the three-step hardy cell screening process. The cells that pass the in vitro viability and transfectability screen proceed to the in vitro ECT capsule viability screen, and the optimal combination of the cell-ECM-scaffold that passed the in vitro capsule viability screen proceed to the in vivo ECT capsule viability screen. Abbreviations aCSF, artificial CSF ECM, extracellular matrix.

In vitro ECT capsule viability screen. For hardy cell candidates that passed the initial cell screen and transfectability screen, the cells were encapsulated and their viability was evaluated using different combinations of extracellular matrix (ECM) and scaffold. The encapsulated performance was examined under optimal tissue culture conditions or under stringent tissue culture conditions (such as artificial CSF). The best... 

Phi-Van, L., von Kries, J.P., Ostertag, W, and Stratling, W. H. (1990) The chicken lysozyme 5 matrix attachment region increases transcription from a heterologous promoter in heterologous cells and dampens position ef fects on the expression of transfected genes. Mol. Cell. Biol. 10 2302-2307. 

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