Available transfection technologies are broadly classified into three groups: chemical, physical, and biological. No one transfection method can be applied to all cell types and experiments

Therefore, the ideal transfection approach should be selected based on your cell type and experimental needs. Specifically, the ability to deliver molecules into cells varies by payload size and cell type. For example, primary and stem cells are typically harder to transfect than more common cell lines. In addition, the method of transfection used should have high efficiency, low toxicity, and minimal effects on normal cell physiology, all while being easy to use and reproducible [1].


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Selecting the right method of transfection

Explore transfection methods and select the method that is best for your cells and experimental application.

Table 1. Important factors for selecting the right transfection method

Selection criteriaCationic lipid-based
Chemical method
Electroporation
Physical method
Viral delivery
Biological method
Efficiency: easy-to-transfect cells+++++++++
Efficiency: hard-to-transfect cells++++++++
Cell viability++++++++
Delivery of large payload (>7 kb)+++++++
High-throughput suitability++++++++
Ease of use+++++++
Biosafety+++++++
Cost per reaction++++++

+++ Excellent for most applications; ++ Good for some applications; + Least recommended, but may be appropriate for some applications.



Chemical gene delivery

Chemical gene delivery methods use carrier molecules that neutralize or impart a positive charge onto nucleic acids.

Cationic lipid transfection

Cationic lipid transfection is one of the most popular methods and can yield high transfection efficiencies in a wide variety of applications and cell types. In addition, this method is versatile, with reagents available for delivery of DNA, RNA, or protein. Specifically, these reagents spontaneously form nucleic acid-cationic lipid reagent complexes that are taken up by the cell via endocytosis.

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Calcium phosphate precipitation

Calcium phosphate precipitation is an easily available and inexpensive transfection method that can be used in many cell types. In this method, calcium phosphate facilitates binding of DNA to the cell surface for uptake via endocytosis.

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DEAE-dextran transfection

DEAE-dextran transfection, one of the earliest chemical transfection methods, is relatively simple to perform and low in cost. In this method, the DEAE-dextran molecule forms a positively charged complex with the nucleic acid that can bind the cell membrane and enter via endocytosis or osmotic shock.

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Cationic polymer transfection

Cationic polymers, which may vary in their degrees of transfection efficiency, are completely water soluble and work by allowing the formation of nucleic acid-polymer complexes. These complexes can adhere to the cell membrane for uptake via endocytosis.

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Physical gene delivery

Physical gene delivery methods allow nucleic acids to be delivered directly into the cytoplasm or nucleus of the cell without the use of chemical carrier molecules.

Electroporation

Electroporation is a popular physical method of transfection that uses an electrical pulse to create temporary pores in cell membranes through which nucleic acids can pass. This method can be used for the rapid transfection of a large number of cells and is applicable in a range of settings, including clinical studies.

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Other physical delivery methods

Other physical gene delivery methods differ from electroporation but still facilitate the direct transfer of nucleic acids into cells without using carrier molecules. These include biolistic particle delivery, in which nucleic acid-coated particles are projected into cells, microinjection, in which a needle is used to directly inject nucleic acids into cells, and laser-mediated transfection, where a laser pulse creates cellular pores.

Explore other physical delivery methods



Biological gene delivery

Biological transfection methods utilize genetically engineered viruses to transfer nucleic acids into cells.

Viral transfection

In viral-mediated transfection, also referred to as transduction, viruses serve as vectors, carrying genes into eukaryotic cells. This method is often used in hard-to-transfect cell types not amenable to other transfection methods and is commonly used in clinical research.

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Related products for transfection methods


References

For Research Use Only. Not for use in diagnostic procedures.

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