Long story short, I am looking to do a stimulation assay in rescued knockout keratinocytes using BMP. Unfortunately for me I am running into issues with low transfection efficiency using lipofectamine on these cells and need a method that would work with a decent efficiency and not cause my cells to differentiate to examine the impacts of the repressed mutant protein as well as some tagged relevant pathway proteins so that their molecular economy can be assessed. I wanted to use viromer red but it does not seem available anywhere in the US. Does any one have advice/ a protocol that worked previously? Have any of you used Viromer Red in the US? If so, how /where did you purchase it?
Hi..
Viromer Red is a cationic liposome-based transfection reagent that has been shown to have high transfection efficiency in a variety of cell types, including keratinocytes. If Viromer Red is not available in the US, you may need to consider using alternative transfection reagents that are suitable for use with keratinocytes.
There are many transfection reagents available that can be used to deliver DNA or RNA to keratinocytes with varying levels of efficiency. Some options to consider include:
Lipofectamine: Lipofectamine is a popular transfection reagent that can be used to deliver DNA or RNA to a variety of cell types, including keratinocytes. However, as you mentioned, transfection efficiency can be low in some cell types, and the reagent can also cause cell differentiation.
Fugene: Fugene is another transfection reagent that can be used to deliver DNA or RNA to a variety of cell types, including keratinocytes. It has a higher transfection efficiency than Lipofectamine in some cell types, but it can also cause cell differentiation.
Oligofectamine: Oligofectamine is a transfection reagent that is specifically designed for transfecting small RNA molecules, such as siRNA and miRNA. It has a lower transfection efficiency than Fugene or Lipofectamine, but it is less toxic to cells and is less likely to cause cell differentiation.
It's worth noting that the optimal transfection reagent and protocol will depend on the specific cell type and transfection conditions, as well as the specific goals of the assay. If you are having difficulty finding a suitable transfection reagent, I recommend consulting a scientist or expert in the field for guidance.
regards
- Badges
- Science topic
- Similar topics
- Biological Science
- Genetics
- Gene Expression