[36] Transient and Stable Transfection Flashcards
What is transfection in molecular biology?
The process of deliberately introducing naked or purified nucleic acids into eukaryotic cells.
What is transient transfection?
A method where introduced DNA exists in cells temporarily and does not integrate into the genome.
What is stable transfection?
A method where introduced DNA is integrated into the genome, providing long-term expression of the transferred gene.
How does the outcome of transient transfection differ from stable transfection?
In transient transfection, gene expression is usually high but short-lived. In stable transfection, gene expression is sustained over a longer period.
What is the main purpose of transfection in research?
To study gene function and regulation, protein production, cell behavior, and more.
What are common methods for transfection?
- Lipofection
- Electroporation
- Microinjection
What is lipofection?
A technique that uses lipid-based reagents to deliver DNA into cells.
What is electroporation?
A technique that uses an electric pulse to introduce DNA into cells.
What is microinjection?
A technique where DNA is injected directly into the nucleus of the cell using a microscopic needle.
What factors can influence the success of transfection?
- Cell type
- Quality and amount of DNA
- Transfection method
What is a selectable marker in the context of stable transfection?
A gene introduced into cells to allow for the identification and selection of cells that have been successfully transfected.
What are examples of selectable markers?
Antibiotic resistance genes, such as neomycin resistance (NeoR) gene.
What is NeoR?
Neomycin resistance gene, a selectable marker often used in stable transfection.
How can the efficiency of transfection be assessed?
By using a reporter gene, such as green fluorescent protein (GFP), that provides a visual indicator of successful transfection.
What is GFP?
Green Fluorescent Protein, a reporter gene that fluoresces green under blue light.
