Tuesday, January 11, 2011

How the compacted DNA gets Attached to the Cell Surface


If a foreign DNA sequence is to be introduced into a cell, it is obviously necessary that the two meet, i.e., that the compacted DNA somehow attaches to the cell surface within and for a reasonable amount of time.


Cell membranes consist of a lipid bilayer into which a number of complex (glyco)protein molecules are inserted or anchored. The dominant mechanism for interaction between the DNA complex and the negatively charged cell surface are electrostatic forces. The negative surface charge is in many cases provided by proteoglycan molecules carrying anionic sulfate groups, which are present on the surface of many cell types. Positively charged complexes may attach themselves to the cell surface via these molecules. The importance of this type of interaction to the success of a transfection has been demonstrated by the following experiment. It has been shown, that DNA charged cationic liposomes fail to transfect so-called Raji cells, which are proteoglycan negative, but transfect genetically modified, proteoglycan positive (syndecan-1), cells of the same cell line with good efficiency.

Electrostatic interaction with the proteoglycans, however, is not the only possibility for interaction between a DNA-carrying transfection agent and a cell surface. Many membrane proteins expose binding sites (receptors) for certain biochemical messenger molecules (ligands).

In general, such receptor proteins control the specific uptake of molecules and make the cell sensitive to hormones and other signal molecules. This natural mechanism can be subverted for DNA transfer. The receptor ligands can be used to increase transfection efficiency in general or they can be used to target the transfection complex to a specific cell or tissue type by evoking an interaction between the transfection complex and a cell-specific receptor. Targeting can, for example, be achieved by introducing ligands such as, insulin, transferrin, lactose, galactose, mannose, folate, poly(acrylic acid) or specific monoclonal antibodies or antibody fragments into the transfection complex. This addition has been shown to dramatically increase the efficiency of transfections with agents such as poly(lysine) or poly(ethyleneimine) for certain cell lines, which otherwise were difficult to transfect. It seems that the improvement is due to the ligand’s ability to subsequently induce receptor-mediated uptake of the DNA into the cell (endocytosis, see below). In addition, receptor mediated transfection can be blocked (controlled) if necessary by complementing the cell culture medium during the transfection with an excess of the free ligand.

Tags: Bio Technology, Bio Genetics, Gene Compaction

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1 comments:

stephen jones on January 11, 2011 at 9:17 PM said...

This is really interesting take on the concept. I never thought of it that way. I came across this site recently which I think it will be a great use of new ideas and informations about DNA. Thanks a lot...

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