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Highly Efficient Transfection of Human THP-1 Macrophages by Nucleofection
Published on: September 2, 2014
High efficiency gene transfer by multiple transfection protocol.
M Yamamoto1, S Okumura, C Schwencke
1Cardiovascular & Pulmonary Research Institute, Allegheny University of the Health Sciences, Pittsburgh, PA 15212, USA.
The Histochemical Journal
|August 14, 1999
Summary
Repeating transfection with LipofectAMINE significantly boosts efficiency. This enhanced protocol doubles transfection rates for the beta-galactosidase gene in 293T cells, improving molecular biology experiments.
Area of Science:
- Molecular Biology
- Cell Biology
- Biotechnology
Background:
- Transfection is crucial for introducing genetic material into cells.
- Conventional methods using polycationic lipids like LipofectAMINE often yield suboptimal efficiencies.
- Improving transfection efficiency is essential for various molecular biology applications.
Purpose of the Study:
- To develop a high-efficiency transfection protocol.
- To enhance the delivery of plasmid DNA into 293T cells using a common polycationic lipid reagent.
Main Methods:
- Utilized LipofectAMINE, a widely available polycationic lipid transfection reagent.
- Transfected 293T cells with a plasmid encoding the beta-galactosidase (beta-gal) gene.
- Employed a repeated transfection strategy, applying the reagent multiple times.
Main Results:
- Conventional transfection achieved less than 40% efficiency.
- The repeated transfection protocol resulted in approximately 80% transfection efficiency.
- Demonstrated a significant, twofold increase in transfection efficiency.
Conclusions:
- Repeating transfection with common polycationic lipids dramatically increases efficiency.
- This simple, optimized protocol enhances gene delivery in cell lines.
- The method offers a valuable improvement for molecular biological experiments requiring efficient transfection.
