Peg-mediated Transfection
Peg-mediated Transfection - Related Videos
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Efficient Polyethylene Glycol (PEG) Mediated Transformation of the Moss Physcomitrella patens
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A simple and efficient method to transform Physcomitrella pantens protoplasts is described. This method is adapted from protocols for Physocmitrella protonemal protoplast and Arabidopsis mesophyll protoplast transformation1.
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Electrophysiology of Scorpion Peg Sensilla
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An Introduction to Transfection
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Transfection is the process of inserting genetic material, such as DNA and double stranded RNA, into mammalian cells. The insertion of DNA into a cell enables the expression, or production, of proteins using the cells own machinery, whereas insertion of RNA into a cell is used to down-regulate the production of a specific protein by stopping translation. While the site of action for transfected RNA is the cytoplasm, DNA must be transported to the nucleus for effective transfection. There, the...
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Source: Van Lidth de Jeude, et al. A Protocol for Lentiviral Transduction and Downstream Analysis of Intestinal Organoids. J. Vis. Exp. (2015).This video demonstrates the process of producing viral particles through transfection of embryonic kidney cells with plasmid–polyethylenimine complexes, which leads to viral genome expression, particle assembly, and virus collection in the medium.
06:41
A Peg Plate Biofilm Assay for Screening Antibacterial Agents
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FAQs
The key mechanistic role of PEG is to alter physical interactions between exogenous nucleic acids and the cell surface. This change can promote closer or more favorable contact during treatment, increasing the opportunity for plasmid DNA and related molecules to cross the membrane. PEG therefore supports uptake through changes in surface and membrane behavior rather than through nucleic acid modification.
Membrane perturbation can help nucleic acids cross the cell membrane, but excessive or prolonged treatment can injure cells. Conditions must therefore balance improved access to the cytoplasm with preservation of cell integrity. Controlling PEG exposure and allowing suitable recovery helps limit treatment-associated damage while retaining the intended genetic delivery effect.
The main variables are PEG concentration, exposure time, cell type, and recovery conditions. These factors influence both how effectively nucleic acids enter cells and how well cells tolerate treatment. A condition that works for one cell type may not produce the same result elsewhere, so transfection efficiency must be considered together with cellular recovery.
A basic workflow exposes cells to PEG in the presence of the selected exogenous nucleic acid, maintains the treatment for a controlled period, and then provides recovery under appropriate conditions. The treatment must be adjusted to the cell type and monitored for excessive damage. Subsequent analysis can determine whether the introduced material supports the intended genetic study.
This method is useful when researchers need transient gene expression, genetic manipulation, or functional testing in cells that are otherwise difficult to transfect. It can provide a way to introduce plasmid DNA or other exogenous nucleic acids without requiring permanent genetic change, making it suitable for experiments that examine gene activity over a limited period.
After introducing exogenous nucleic acids, researchers can examine transient gene expression and use the resulting genetic manipulation to study gene function. The method is especially relevant when cellular responses need to be assessed after delivery into the cytoplasm. Interpretation should account for treatment conditions, because PEG concentration, exposure time, cell type, and recovery can affect the outcome.