APR January/February 2022 - 33

« DRUG DEVELOPMENT
Figure 5. The mRNA Transfection Process4
Although viral delivery methods are highly efficient, they can be quite
laborious. Most viruses require containment and careful monitoring
of biosafety levels. Before performing viral transfections, it is also
important to consider several limiting factors such as the lytic nature
of viral vectors, cell line packaging and host-cell specificity.
mRNA Transfection Process
The process of mRNA transfection is simpler: 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 (Figure 5). The transient nature of mRNA transfection is desirable
for a number of applications, including cellular
genome editing (CRISPR/Cas9) and vaccines.
reprogramming,
Advantage of mRNA Transfection
mRNA transfection has some advantages over DNA transfection,
which are especially helpful 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 repetition.
* 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.
www.americanpharmaceuticalreview.com |
| 33
Figure 6. Transduction, what happens within the cells
»
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APR January/February 2022

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