Universal Transfection Reagents: Improved Efficiency and Decreased Cellular Toxicity

Fluorescent microscopy images of cells after being transfected.

TTransfection alters the genetic makeup of eukaryotic cells by introducing foreign nucleic acids, including DNA, RNA, and small non-coding RNAs such as siRNA, shRNA, and miRNA. Scientists use transfection techniques to advance cell research and improve drug discovery by allowing researchers to characterize cellular processes and study the molecular mechanisms of disease.1–3

Planning for a successful transfection

Researchers deliver transfected nucleic acids as oligonucleotides or in a viral vector or plasmid, which carries the genetic material to host cells. The transfection can be stable or transient. Stable transfection results in long-term sustained expression, whereas expression will ultimately be lost after transient transfection as host cells replicate. Scientists apply stable transfection for large-scale, long-term genetic and pharmacological studies. Transient transfection is useful for short-term studies, such as investigating the effects of gene activation or deactivation.two

When planning transfection experiments, researchers must first select a delivery method to transfer nucleic acids to target cells. The delivery method can be physical, such as electroporation, or chemical involving lipid-based or non-lipid-based reagents. Delivery methods affect the host cell surface and facilitate entry of nucleic acid into the cell. In the case of chemical delivery methods, the reagent forms a complex with the nucleic acid to enhance contact with the cell membrane. The ability to successfully transfect a cell varies depending on the cell type, the protocol, and the composition of the transfection reagent.1.2

Transfection challenges

Four blue containers of Roche® X-tremeGENE™ Transfection Reagents.

Transfection often causes cytotoxicity due to the effects of the transfection reagents on the cell surface, which can stress the cells. In addition, transfection reagents can be expensive and have very specific applications, so researchers often need to purchase different reagents depending on the nucleic acids and cell types they transfect. When choosing transfection reagents, researchers must identify the cell type and culture conditions for their experiment. Rare cell cultures, such as neurons or primary cells, require reagents that facilitate delivery of nucleic acids into cells that are difficult to transfect. Investigators should also consider the amount of reagent they will need before selecting an appropriate transfection reagent, as this will affect cost and toxicity. The ideal product will minimize the number of different reagents a researcher needs and optimize the effectiveness of their experiments. Therefore, an ideal reagent has multiple applications, low cytotoxicity, and high transfection efficiency.1–3

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Roche X-tremeGENE™ Transfection Reagents® efficiently transfect many cell types, from common to rare and primary cells. Scientists can choose from a variety of X-tremeGENE™ transfection reagents based on their experimental needs, to deliver a variety of molecules in different applications, including lentiviral production, gene knockout, and gene editing.1.4–7

An all-in-one solution

Researchers looking for an all-in-one reagent to use for different experiments can choose the new X-tremeGENE™ 360 Transfection Reagent, a reliable, versatile, high-throughput solution for delivering a variety of nucleic acids into many different cell types. This innovative reagent forms a complex with DNA or RNA and can transfect siRNA/miRNA, plasmid DNA, and CRISPR/Cas9 materials into animal or insect cells with high efficiency. It is a universal polymer designed for a wide range of eukaryotic cells, including many cell lines that do not transfect well with other reagents. The X-tremeGENE™ 360 Reagent works well in the presence or absence of serum and can be used by researchers for transient transfection, stable transfection, siRNA expression, and CRISPR gene editing. Finally, the X-tremeGENE™ 360 reagent is cost effective and cost effective. 1 mL of X-tremeGENE™ 360 Transfection Reagent can be used to perform up to 10,000 transfections in 96-well plates. Because it produces minimal changes in cytotoxicity and cell morphology when adequate numbers of cells are transfected, it eliminates the need to change media after adding the transfection reagent, saving time and media expense.1.6 This universal and efficient transfection reagent with minimal cytotoxicity is an attractive solution for researchers looking to streamline their transfection protocols and declutter their cell culture room freezers.

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  1. “X-tremeGENE™ Transfection Reagent Comparison Guide,” https://www.sigmaaldrich.com/CA/en/technical-documents/technical-article/genomics/advanced-gene-editing/general-recommendation-for- transfection-reagent-selection, accessed 4 September 2022.
  2. ZX Chong et al., “Transfection Types, Methods, and Strategies, A Technical Review” Peer J9:1-37, 2021.
  3. “Introduction to cell transfection”, https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/cell-culture-and-cell-culture-analysis/transfection-and-gene-editing/ transfection-reagents, accessed 4 September 2022.
  4. “Lentiviral Production Using X-tremeGENE HP Transfection Reagent,” https://www.sigmaaldrich.com/US/en/technical-documents/protocol/cell-culture-and-cell-culture-analysis/transfection-and- gene-editing/xtghp-lenti-protocol, accessed 4 September 2022.
  5. “X-tremeGENE™ HP DNA Transfection Reagent,” https://www.sigmaaldrich.com/US/en/product/roche/xtghpro, accessed September 4, 2022.
  6. “X-tremeGENE™ 360 Transfection Reagent,” https://www.sigmaaldrich.com/US/en/product/roche/xtg360ro, accessed September 4, 2022.
  7. “X-tremeGENE™ 9 DNA Transfection Reagent,” https://www.sigmaaldrich.com/US/en/product/roche/xtg9ro, accessed September 4, 2022.

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