Lipofection

Principle

Lipofection is a lipid-based transfection technology which belongs to biochemical methods including also polymers, DEAE dextran and calcium phosphate.
Lipofection principle is to associate nucleic acids with cationic lipid formulation. The resulting molecular complexes, known as lipoplexes, are then taken up by the cells.
The main advantages of lipofection are its high efficiency, its ability to transfect all types of nucleic acids in a wide range of cell types, its ease of use, reproducibility and low toxicity. In addition this method is suitable for all transfection applications (transient, stable, co-transfection, reverse, sequential or multiple transfections…), high throughput screening assay and has also shown good efficiency in some in vivo models.

How does it work?

The lipid-based reagents used for lipofection are generally composed of synthetic cationic lipids that are often mixed with helper lipids such as DOPE (1,2-dioleoyl-phosphatidyl-ethanolamine) or cholesterol. These lipids mixture assembles in liposomes or micelles with an overall positive charge at physiological pH and are able to form complexes (lipoplexes) with negatively charged nucleic acids through electrostatics interactions. The association of the lipid-based transfection reagent with nucleic acids results in a tight compaction and protection of the nucleic acids and these cationic complexes are mainly internalized by endocytosis.
Once inside the cells two mechanisms leading to the nucleic acids release into the cytoplasm have been described. One relies on the endosomes buffering capacity of the polycationic residues (called “proton sponge effect”). The other describes the ability of cellular negatively charged lipids to neutralize the cationic residues of the transfection reagent leading to destabilization of endosomal membranes.
Finally, the cellular and molecular events leading to the nuclear uptake of DNA (not required for siRNA) following by gene expression remain highly speculative. However, the significance of cell division on transfection efficiency favours the assumption that nuclear membrane disruption during the mitosis process promote DNA nuclear uptake. Nonetheless, transfections of primary cells (non-dividing) and in vivo are also achievable with lipofection demonstrating that DNA can make its way to the nucleus where gene expression takes place.

Tee-Technology

The cationic lipids (lipoplexes) and polymers (polyplexes) are the most employed non-viral gene delivery systems. The Tee-Technology (Triggered Endosomal Escape) combines and exploits the properties of both entities to achieve extremely efficient nucleic acids delivery into cells. Indeed, this new generation of lipopolyamines contains a lipophilic part, such as lipids, and a charged polyamine moiety, such as cationic polymers. These moieties act in synergy to ensure a tight nucleic acids compaction and protection and a very efficient destabilization of the endosomal membrane which allows the release of large nucleic acids amounts in the cytosol and DNA nuclear uptake. A particular focus on the synthesis of fully biodegradable entities was integrated. In this way, the transfection reagents do not interfere with cellular mechanisms, high cell viability is maintained in every experiment and any potential secondary effects are avoided.

What are the applications?

Transfection efficiency combined with high transgene expression level or high gene silencing and minimized cytotoxicity depends on multiple critical parameters. Those factors include cell type, plasmid DNA characteristics (size, promoter, reporter gene) & purity, siRNA sequence & purity, cell culture conditions (medium with or without serum, cell number, absence of contaminations…), amount of nucleic acids and reagents, transgene assays to name a few. Consequently, transfection reagents need to be specifically designed according to the nucleic acids to be delivered (DNA, siRNA, mRNA, ODN, shRNA etc.) and the cell types used in order to achieve optimal efficiency. In this context, OZ Biosciences has developed several outstanding transfection reagents:Lipofection method is especially suitable for immortalized cells.
Please contact our technical support: techsupport(at)bio-connect.nl for the list of cells successfully tested.

The major Tee-Technology advantages are:

  • Compaction of DNA in nanoparticles efficiently internalized by cells
  • Protection of nucleic acids against nucleases degradation
  • Efficient membrane destabilization and DNA delivery
  • Highly efficient even with low amounts of nucleic acids
  • Biodegradability

How do I use Lipofection reagents?

The protocol is a very straightforward and easy procedure:
  1. Prepare the DNA and the Reagent solutions.
  2. Mix them together and incubate 20 min.
  3. Add to your cells.

Transfection Reagents

Cat. No.
DescriptionAmount

DreamFect™ Gold Transfection Reagent

DG80500
DreamFect Gold 500 Transfection Reagent
0,5 ml
DG81000
DreamFect Gold 1000 Transfection Reagent
1 ml
DG85000
DreamFect Gold 5000 Transfection Reagent
5 x 1 ml

DreamFect™ Transfection Reagent

DF40500
DreamFect 500 Transfection Reagent
0,5 ml
DF41000
DreamFect 1000 Transfection Reagent
1 ml
DF45000
DreamFect 5000 Transfection Reagent
5 x 1 ml

Lullaby™ Transfection Reagent

LL70500
Lullaby 500 Transfection Reagent
500 ul
LL71000
Lullaby 1000 Transfection Reagent
1000 ul
LL73000
Lullaby 3x1ml Transfection Reagent
3 x 1 ml

VeroFect™ Transfection Reagent

VF60250
VeroFect 250 Transfection Reagent
250 ul
VF60500
VeroFect 500 Transfection Reagent
500 ul
VF61000
VeroFect 1000 Transfection Reagent
1 ml
VF65000
VeroFect 5000 Transfection Reagent
5 x 1 ml

FlyFectin™ Transfection Reagent

FF50500
FlyFectin 500 Transfection Reagent
500 ul
FF51000
FlyFectin 1000 Transfection Reagent
1 ml
FF55000
FlyFectin 5000 Transfection Reagent
5 x 1 ml

EcoTransfect™ Transfection Reagent

ET10500
EcoTransfect 500 Transfection Reagent
0,5 ml
ET11000
EcoTransfect 1000 Transfection Reagent
1 ml
ET13000
EcoTransfect 3000 Transfection Reagent
3 x 1 ml

COSFect Transfection Reagent

CF10500
COSFect 500 Transfection Reagent
500 ul
CF11000
COSFect 1000 Transfection Reagent
1 ml
CF12500
COSFect 5x1000 Transfection Reagent
5 x 1 ml

HeLaFect Transfection Reagent

HF20500
HeLaFect 500 Transfection Reagent
500 ul
HF21000
HeLaFect 1000 Transfection Reagent
1 ml
HF25000
HeLaFect 5x1000 Transfection Reagent
5 x 1 ml

Ab-DeliverIN™ Transfection Reagent

AI20100
Ab-DeliverIN 100 Transfection Reagent+ FITC
100 ul
AI20250
Ab-DeliverIN 250 Transfection Reagent+ FITC
250 ul
AI20500
Ab-DeliverIN 500 Transfection Reagent+ FITC
500 ul
AI21000
Ab-DeliverIN 1000 Transfection Reagent+ FITC
1 ml

Pro-DeliverIN™ Transfection Reagent

PI10100
Pro-DeliverIN 100 Transfection Reagent+ R-Phycoerythrin
100 ul
PI10250
Pro-DeliverIN 250 Transfection Reagent+ R-Phycoerythrin
250 ul
PI10500
Pro-DeliverIN 500 Transfection Reagent+ R-Phycoerythrin
500 ul
PI11000
Pro-DeliverIN 1000 Transfection Reagent+ R-Phycoerythrin
1 ml



Source:
Website: OZ Biosciences

Related to:
Brands: OZ Biosciences
Product groups: Molecular Biology