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.

Step 1 : Mix nucleic acid with DEAE

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.

Step 2 : Add the nucleic acid-DEAE to cells

Add the nucleic acid-DEAE-dextran complexes to the cells, which adhere to the cell surface via electrostatic interactions.

Step 3 : Induce the uptake

Induce the uptake of the nucleic acid-DEAE-dextran complexes by osmotic shock using DMSO or glycerol.

Step 4 : Wash cells

Wash cells to remove the complexes and incubate to allow gene expression.

Step 5 : Assay cells

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.

References

  1. Pagano JS, Vaheri A. Enhancement of infectivity of poliovirus RNA with diethylaminoethyl-dextran (DEAE-D). Arch Gesamte Virusforsch. 1965;17(3):456–464. doi: 10.1007/BF01241201. PMID: 4286824.

For Research Use Only. Not for use in diagnostic procedures.