Gene editing focuses on the effect of gene modification on loss or gain of cell functions, often studied by insertion of a particular (modified) gene through plasmid transfection. Unfortunately, a wide variety of cells are difficult to transfect with plasmid DNA, including adherent cells (neurons or primary cells), suspension cells (lymphocytes) and various cancer cell lines. This crucial step in gene expression experiments can limit you enormously in progression of your research.
What if you could tackle gene expression difficulties with messenger RNA (mRNA) transfection?
DNA transfection versus mRNA transfection: the principles
The DNA transfection process
To express a (modified) gene of interest in your cells, the most well-known and used method is DNA transfection. To this end, a plasmid DNA will form a complex with the transfection reagent and will attach to the cell membrane. Through endocytosis this reagent-DNA complex is taken up by the cell and the DNA is released into the cytoplasm. It will need to enter the nucleus before it can be transcribed, via pre-mRNA, into mRNA. This entrance of the DNA into the nucleus is a crucial step, since mRNA is needed for translation of the gene into a protein, but also the most difficult step to achieve with hard-to-transfect cells.
Once mRNA is formed, the mRNA molecule will be released into the cytoplasm and translated into a protein.
The mRNA transfection process
The process of mRNA transfection is more simple: mRNA is directly delivered and expressed in the cytoplasm and thus does not require to cross the nuclear membrane. After delivery, it can immediately be translated into a protein in the cytoplasm.
Since this gene expression method does not require the mRNA molecule to enter the nucleus, this is an ideal solution for transfection of stem cells, neurons, lymphoma and many more hard-to-transfect cell types (see Download section below). The transient nature of mRNA transfection is desirable for a number of applications, including cellular reprogramming, genome editing (CRISPR/Cas9) and vaccines.
Advantages of mRNA transfection
mRNA transfection has some advantages over DNA transfection, which are especially helpfull when working with difficult cell types in gene editing research:
- mRNA tends to transfect more efficiently with higher efficiencies reached (>80%) for hard-to-transfect cell types
- mRNA transfection results in more rapid protein expression, as it doesn’t need to be transcribed first
- protein expression is easily adjustable by changing mRNA concentrations or transfection repitition
- mRNA transfection does not require nucleus entry and is therefore perfectly suited for non-dividing cells
- no risk of insertional mutagenesis, hence no genome modification of the transfected cell
mRNA transfection with jetMESSENGER
The jetMESSENGER mRNA transfection reagent has proven benefits over other mRNA transfection reagents:
- Unmatched transfection efficiency
- High efficiency on a wide variety of difficult to transfect cells including neuronal cells, immune cells, stem cells and cancer cell lines
- Extremely gentle on cells
- Easy to scale up and down
- 2-fold lower costs*
*compared to Lipofectamine™ MessengerMAX™