JTransfection changes the genetic makeup of eukaryotic cells by introducing foreign nucleic acids including DNA, RNA, and small non-coding RNAs such as siRNAs, shRNAs, and miRNAs. Scientists use transfection techniques to advance cellular research and improve drug discovery by enabling researchers to characterize cellular processes and study the molecular mechanisms of disease.1–3
Planning a successful transfection
Researchers deliver transfected nucleic acids as oligonucleotides or in a viral or plasmid vector, which carries the genetic material into host cells. Transfection can be stable or transient. Stable transfection results in long-term sustained expression, while expression will eventually be lost after transient transfection as host cells replicate. Scientists apply stable transfection for long-term, large-scale genetic and pharmacological studies. Transient transfection is useful for short-term studies, such as studying the effects of knocking or knocking out the gene.2
When planning transfection experiments, researchers must first select a delivery method to transfer nucleic acids into target cells. The method of administration can be physical, such as electroporation, or chemical involving lipid-based or non-lipid-based reagents. Methods of administration affect the surface of the host cell and facilitate entry of the nucleic acid into the cell. In the case of chemical delivery methods, the reagent forms a complex with the nucleic acid to improve contact with the cell membrane. The ability to successfully transfect a cell varies by cell type, protocol, and composition of the transfection reagent.1.2
Transfection often causes cytotoxicity due to the effects of transfection reagents on the cell surface, which can stress cells. Additionally, 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 are transfecting. When choosing transfection reagents, researchers should identify the cell type and culture conditions for their experiment. Cultures of rare cells such as neurons or primary cells require reagents that facilitate nucleic acid delivery into difficult-to-transfect cells. Researchers 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 maximize the efficiency of their experiments. Therefore, an ideal reagent has multiple applications, low cytotoxicity and high transfection efficiency.1–3
X-tremeGENE™ Roche transfection reagents® effectively transfect many cell types, from common cells to rare and primary cells. Scientists can choose from a range of X-tremeGENE™ transfection reagents depending on their experimental needs, to deliver a variety of molecules in different applications, including lentiviral production, gene silencing and gene editing.1.4–7
An all-in-one solution
Researchers looking for an all-in-one reagent to use in different experiments can choose the new X-tremeGENE™ 360 Transfection Reagent, a powerful, versatile and reliable 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 mistransfected by other reagents. The X-tremeGENE™ 360 reagent works well in the presence or absence of serum and researchers can use it for transient transfection, stable transfection, siRNA expression and CRISPR gene editing. Finally, the X-tremeGENE™ 360 reagent is economical and fast. 1ml of X-tremeGENE™ 360 Transfection Reagent can be used to perform up to 10,000 transfections in 96-well plates. Because it produces minimal cytotoxicity and cell morphology changes when an adequate number of cells are transfected, it eliminates the need to change media after adding the transfection reagent, saving time and media spending.1.6 This universal and efficient transfection reagent with minimal cytotoxicity is an attractive solution for researchers looking to optimize their transfection protocols and declutter their cell culture room freezers.
- “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-selectionaccessed September 4, 2022.
- ZX Chong et al., “Types, methods and strategies of transfection, a technical review”, PeerJ9:1-37, 2021.
- “Introduction to Cellular Transfection”, https://www.sigmaaldrich.com/US/en/technical-documents/technical-article/cell-culture-and-cell-culture-analysis/transfection-and-gene-editing/transfection-reagentsaccessed September 4, 2022.
- “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-protocolaccessed September 4, 2022.
- “X-tremeGENE™ HP DNA Transfection Reagent”, https://www.sigmaaldrich.com/US/en/product/roche/xtghproaccessed September 4, 2022.
- “X-tremeGENE™ 360 Transfection Reagent”, https://www.sigmaaldrich.com/US/en/product/roche/xtg360roaccessed September 4, 2022.
- “X-tremeGENE™ 9 DNA Transfection Reagent”, https://www.sigmaaldrich.com/US/en/product/roche/xtg9roaccessed September 4, 2022.