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Related Experiment Videos

Rapid transformant selection by human complement using HRF20 (CD59) cDNA as a selection marker

H Takizawa1, K Takahashi, T Murakami

  • 1Department of Molecular Biology, Nagoya City University School of Medicine, Japan.

European Journal of Immunology
|October 1, 1993
PubMed
Summary
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The homologous restriction factor (HRF20, CD59) gene can make cells resistant to complement attack. This offers a rapid and less damaging method for selecting successfully transfected cells.

Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Biology

Background:

  • The complement system is a crucial part of innate immunity.
  • Membrane attack complexes (MACs) are key mediators of complement-mediated cell lysis.
  • Homologous restriction factor (HRF20, CD59) is a known inhibitor of MAC formation.

Purpose of the Study:

  • To investigate the potential of HRF20 (CD59) as a selectable marker for gene transfection.
  • To evaluate the efficiency and speed of HRF20-mediated complement resistance for cell selection.

Main Methods:

  • Transfection of heterologous cells with HRF20 cDNA.
  • Exposure of transfected cells to human complement.
  • Assessment of cell survival as a measure of transfection efficiency.

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Main Results:

  • Transfection with HRF20 rendered complement-sensitive cells resistant to complement-mediated lysis.
  • This method allowed for the selection of transfected cells within 8 hours.
  • Minimal cell damage was observed due to the short incubation time with complement.

Conclusions:

  • HRF20 (CD59) serves as an effective and rapid selectable marker for gene transfection.
  • Complement resistance mediated by HRF20 offers a less toxic alternative to traditional drug resistance selection methods.