What is DEAE-dextran transfection?
DEAE-dextran is a polycationic derivative of the carbohydrate polymer dextran and was one of the first chemical reagents used to transfer nucleic acids into cultured mammalian cells [1]. In addition, DEAE-dextran has been used to increase the efficiency of lentiviral transduction.
In DEAE-dextran-mediated transfection, the cationic DEAE‑dextran molecule tightly associates with the negatively charged backbone of the nucleic acid. The net positive charge of the resulting nucleic acid-DEAE‑dextran complex allows it to adhere to the cell membrane and enter into the cytoplasm via endocytosis or osmotic shock induced by DMSO or glycerol.
Basic steps for performing DEAE-dextran-mediated transfection
Mix nucleic acid with DEAE-dextran solution in transfection medium or phosphate-buffered saline solution. Nucleic acid-DEAE-dextran complexes are formed via electrostatic interactions between the polymer and phosphate backbone of the nucleic acid.
Add the nucleic acid-DEAE-dextran complexes to the cells, which adhere to the cell surface via electrostatic interactions.
Induce the uptake of the nucleic acid-DEAE-dextran complexes by osmotic shock using DMSO or glycerol.
Wash cells to remove the complexes and incubate to allow gene expression.
Assay cells for transient gene expression.
Mix nucleic acid with DEAE-dextran solution in transfection medium or phosphate-buffered saline solution. Nucleic acid-DEAE-dextran complexes are formed via electrostatic interactions between the polymer and phosphate backbone of the nucleic acid.
Add the nucleic acid-DEAE-dextran complexes to the cells, which adhere to the cell surface via electrostatic interactions.
Induce the uptake of the nucleic acid-DEAE-dextran complexes by osmotic shock using DMSO or glycerol.
Wash cells to remove the complexes and incubate to allow gene expression.
Assay cells for transient gene expression.
Advantages and disadvantages of DEAE-dextran transfection method
DEAE-dextran transfection is advantageous in that it is simple to perform and affordably available; however, this method also has disadvantages, including potential cytotoxicity and low transfection efficiencies. It should also be noted that this method requires reduced-serum media during the transfection procedure.
Advantages
- Relatively simple to perform
- Reproducible results
- Affordable
Disadvantages
- Can be cytotoxic
- Low transfection efficiency for a range of cells (typically less than 10% in primary cells)
- Limited to transient transfection
Visit Transfection Basics to learn more about performing transfection in your lab.
For Research Use Only. Not for use in diagnostic procedures.