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Efficient cell transfection with melamine-based gemini surfactants.

Serena Perrone1, Michele Usai, Paolo Lazzari

  • 1Kosterlitz Centre for Therapeutics, Institute of Medical Sciences, School of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, Scotland, United Kingdom.

Bioconjugate Chemistry
|January 10, 2013
PubMed
Summary

New melamine gemini surfactants offer efficient and low-toxicity DNA delivery for various cell lines, including difficult-to-transfect neurons. These novel reagents outperform established methods like Lipofectamine 2000.

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Area of Science:

  • Biochemistry
  • Materials Science
  • Molecular Biology

Background:

  • Gene delivery is crucial for research and therapeutics.
  • Developing efficient, safe, and versatile transfection reagents remains a challenge.
  • Melamine-based compounds offer unique structural properties for molecular applications.

Purpose of the Study:

  • To synthesize and evaluate novel melamine-based gemini surfactants for DNA transfection.
  • To optimize transfection protocols for different cell lines using these novel surfactants.
  • To compare the efficiency and toxicity of these surfactants against a gold-standard reagent.

Main Methods:

  • Synthesis of gemini surfactants with varying lipophilic chain lengths (C8-C16) based on melamine scaffolds.
  • Transfection of A549, U87 MG, and Bristol 8 cell lines using a maxGFP expressing plasmid.

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  • Optimization of transfection parameters including cell confluence, reagent/DNA ratio, compaction, and time.
  • Assessment of transfection efficiency and cytotoxicity in various cell types, including primary DRG neurons.
  • Evaluation of reagent performance in the presence and absence of serum.

    • Main Results:

    • Successfully synthesized melamine gemini surfactants with tunable lipophilic chain lengths.
    • Demonstrated efficient DNA transfection in multiple cancer cell lines (A549, U87 MG, Bristol 8).
    • Achieved superior transfection efficiency with C12 and C14 surfactants compared to Lipofectamine 2000, with low toxicity.
    • Successfully transfected primary dorsal root ganglion (DRG) neurons, a notoriously difficult cell type.
    • Observed complete inhibition of transfection in the presence of serum.

    Conclusions:

    • Melamine-based gemini surfactants are effective, low-toxicity reagents for DNA transfection.
    • Optimized C12 and C14 surfactants show significant potential for gene delivery applications.
    • These novel surfactants offer advantages in synthesis, stability, and ease of use.
    • Further development could lead to improved therapeutic gene delivery strategies.