Lipid-based transfection, also known as "lipofection" or "liposome-based transfection", is a biochemical method that uses a lipid complex to deliver nucleic acids into cells. Reagents used for lipofection are generally composed of synthetic cationic lipids that are often mixed with helper lipids such as 1,2-dioleoyl-phosphatidyl-ethanolamine (DOPE) or cholesterol. These lipids mixture assembles in liposomes or micelles with an overall positive charge at physiological pH and are able to form complexes (called lipoplexes) with negatively charged nucleic acids through electrostatics interactions. The lipoplexes bind to the cell surface by non-specific, electrostatic interactions with the negatively charged cell surface, and enter cells by endocytosis or endocytosis-like mechanisms. Once inside, the pH of the endosome compartments drops and part of the bound nucleic acids escape from early endosomes into the cytosol. Cytoplasmic transport of endosomes plays an active role in bringing the transfecting material near the perinuclear region. The lipoplexes then dissociate and the released DNA enters the nucleus.
The advantages of lipofection are high efficiency, ease of use, high reproducibility, low toxicity, and the ability to transfect all types of nucleic acids in a wide range of cell types. In addition, the method is suitable for all transfection applications, such as transient, stable, co-transfection, reverse, sequential or multiple transfections, and high-throughput screening assays.
Cationic lipids (lipoplexes) and polymers (polyplexes) are the most widely used non-viral gene delivery systems. The Triggered Endosomal Escape (Tee)-Technology combines the properties of these two delivery systems to achieve extremely efficient delivery of nucleic acids to cells. This new generation of lipopolyamines contains a lipophilic part, such as lipids, and a charged polyamine moiety, such as cationic polymers. These parts act synergistically to ensure a tight nucleic acid compaction and protection and a very efficient destabilization of endosomal membrane, which allows the release of large amounts of nucleic acid in the cytoplasm and DNA nuclear uptake. Being integrated as fully biodegradable entities, the transfection reagents do not interfere with cellular mechanisms, ensuring high cell viability in each experiment and avoiding any potential secondary effects.
Tee-Technology's main advantages are:
Transfection efficiency is related to transgenic expression level, gene silencing and cytotoxicity, which are affected by factors including cell type, plasmid DNA characteristics (size, promoter, reporter gene) and purity, siRNA sequence and purity, cell culture conditions, nucleic acids, reagents, etc.
To achieve optimal efficiency, transfection reagents need to be specifically selected according to the type of nucleic acid to be delivered and the cell type to be transfected. Amerigo Scientific offers high efficiency transfection reagents based on Tee-Technology to meet your assay needs.
|DreamFect Gold||For all nucleic acids, achieving superior transgene expression level|
|DreamFect / DreamFect Stem||For all nucleic acids, for all cells including suspension cell lines|
|Lullaby® / Lullaby® Stem||For siRNA applications|
|RmesFect / RmesFect Stem||For mRNA applications|
|EcoTransfect||For popular cell lines and routine transfection at low cost|
|FlyFectIN||For insect cells transfection|
|HeLaFect||For HeLa cells transfection|
|VeroFect||For Vero cells transfection|
|COSFect||For COS cells transfection|
|Ab-DeliverIN||For intracellular antibody delivery|
|Pro-DeliverIN||For intracellular protein delivery|
The protocol is a very straightforward and easy procedure:
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