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Predicting Transfection Efficiency – Why is it So Important?

Ensuring efficient transfection is essential for gene therapy, CRISPR/Cas9, and many other applications in industry and research laboratories. However, many factors affect transfection efficiency. Furthermore, the methods available to assess this efficiency can be laborious and time-consuming. If you're involved in gene expression and modulation research, you may be familiar with these issues. But as always, science evolves, and new alternatives arise to facilitate our work and accelerate our discoveries.

What is transfection?

For those just starting transfection experiments, let's clarify some basic concepts before delving into the topic.

Transfection is a method that allows inserting foreign nucleic acids (DNA and RNA) into eukaryotic cells, [1]. With this powerful technique, researchers can modify the host cell's genetic makeup. The applications are endless; you can study disease's mechanisms, discover biomarkers for diagnosis and prognosis, or develop gene therapies to treat genetic diseases [1].

Examples of applications of transfection techniques [1]:

  • Gene silencing

  • Stable cell line generation

  • Virus production

  • Large-scale protein production

  • Stem cell reprogramming and differentiation

Types of transfection

There are two types of transfection:

  • Stable – Stable transfection involves integrating the foreign nucleic acid into the host cell nucleus maintaining its expression in the long term, [1].

  • Transient – This type of transfection does not involve integrating nucleic acids into the host cell genome. For instance, you can use a plasmid or oligonucleotides to transfect nucleic acids. However, gene expression will be lost with cell replication, [1].

Briefly, stable transfection is helpful if you need large amounts of protein production. In contrast, transient transfection is more useful to investigate the short-term effects of a gene knock-out or knock-in, [1].

The options for delivering the genetic material into the host cells include vectors like viruses or plasmids, [1]. Common transfection techniques are calcium phosphate precipitation, lipofection, and electroporation, which increase cell permeability through an electrical field, [1].

Why is transfection efficiency important?

If you're a researcher working with cell transfection, you know how challenging it can be to optimize the process.