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Selection of transfected mammalian cells
R Mortensen1, J D Chestnut, J P Hoeffler
1Brigham and Women's Hospital, Boston, Massachusetts, USA.
Current Protocols in Neuroscience
|April 23, 2008
Summary
Creating stable mammalian cell lines for gene function analysis requires selecting successfully transfected cells. This study details methods for determining selection conditions and optimizing transfection using selectable markers.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- Gene function studies often necessitate stable mammalian cell lines with integrated genes.
- Transfection efficiency for stable integration is low, requiring selection methods.
- Dominant selectable markers are crucial for isolating successfully transfected cells.
Purpose of the Study:
- To present procedures for determining selection conditions for stable transfection.
- To discuss commonly used selectable markers for mammalian cell selection.
- To provide protocols for optimizing transfection conditions.
Main Methods:
- Determining selection conditions for stable transfectants.
- Utilizing thirteen common selectable markers for mammalian cell selection.
- Employing a simple staining assay to optimize transfection.
Main Results:
- Established protocols for determining selection conditions and achieving stable transfection.
- Detailed conditions for thirteen widely used mammalian cell selection markers.
- Demonstrated optimization of transfection efficiency through a staining assay.
Conclusions:
- Effective selection strategies are vital for generating stable mammalian cell lines for research.
- A range of selectable markers and optimization protocols are available to facilitate gene function analysis.
- Optimized transfection and selection increase the yield of desired cell lines.
