Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Highly Efficient Gene Delivery into Transfection-Refractory Neuronal and Astroglial Cells Using a Retrovirus-Based Vector

  • Kim, Byung Oh (Department of Applied Biology, College of Life Science and Natural Resources, Sangju National University) ;
  • Pyo, Suhkneung (Division of Immunopharmacology, College of Pharmacy, Sungkyunkwan University)
  • Published : 2005.04.01
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Abstract

Introduction of foreign genes into brain cells, such as neurons and astrocytes, is a powerful approach to study the gene function and regulation in the neuroscience field. Calcium phosphate precipitates have been shown to cause cytotoxicity in some mammalian cells and brain cells, thus leading to low transfection efficiency. Here, we describe a retrovirus-mediated gene delivery method to transduce foreign genes into brain cells. In an attempt to achieve higher gene delivery efficiency in these cells, we made several changes to the original method, including (1) use of a new packaging cell line, Phoenix ampho cells, (2) transfection of pMX retroviral DNA, (3) inclusion of 25 mM chloroquine in the transduction, and (4) 3- 5 h incubation of retroviruses with target cells. The results showed that the modified protocol resulted in a range of 40- 60% gene delivery efficiency in neurons and astrocytes. Furthermore, these results suggest the potential of the retrovirus-mediated gene delivery protocol being modified and adapted for other transfection-refractory cell lines and primary cells.

Keywords

References

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