This video shows how dendritic cells growing in culture are transfected with a plasmid containing a gene for the Green fluorescent protein (GFP).
Transfection is the process of deliberately introducing nucleic acids into eukaryotic cells. Various methods can be used to transfect. Electroporation, shown in this video, is a technique by which an externally applied electrical field applied to the cells makes the plasma membrane of a cell temporarily porous so that molecules such as DNA can pass freely through it.
Once the DNA is inside the cells the genes in this DNA molecule can be “read” by the cell machinery and made into proteins. In this example the molecule DNA entered the cell has information to produce the green fluorescent protein GFP. Therefore the cells that have taken in the DNA can now make GFP and glow green.
This video is inspired in the DC transfection I was doing while I was in graduate school. The results using transfected DCs are published in Figure 5.f in the paper A CD74-dependent MHC class I endolysosomal cross-presentation pathway.
Below are some links with more information on the topics discussed in this post.
- Overview of Transfection and electroporation from Wikipedia
- Product I used to transfect DCs: Nucleofector™ Kits for Mouse Dendritic Cells by Lonza
- Protocol for Generation of Bone Marrow Derived Murine Dendritic Cells by Journal of Visualized Experiments
- Products to isolate dendritic cells from mice
- Overview of Dendritic cells from Wikipedia
- Overview of GFP from Wikipedia