Transfection, Transduction, and Membrane Fusion

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The term “transduction” is used to describe a virus-mediated DNA transfer into cells. In contrast to transfection of cells with foreign DNA or RNA, no transfection reagent is needed here. The viral vector, itself, also called virion, is able to infect cells and transport the DNA directly into the nucleus, independent of further actions. After the release of the DNA into the nucleus, the protein of interest is produced using the cells’ own machineries.

Adenoviral vectors have proven to be a very successful tool in many different human and rodent cell types. Transduction efficiencies of up to 100% can easily be achieved. Moreover, this method also gives access to difficult-to-transfect cells, such as primary cells.

With the rAV-LifeAct adenoviral vectors, ibidi provides a perfect tool for visualizing F-actin in difficult-to-access cells (e.g., endothelial cells) without interfering with the cytoskeletal dynamics.

Transduction of mammalian cells using an adenoviral vector (here rAV-LifeAct)

Recombinant lentiviral vectors have been shown to be powerful tools for stable gene transfer to both dividing and non-dividing cells in vitro and in vivo as they integrate into the host genome. They have a broad host cell range that also includes cell types such as neurons, lymphocytes, and macrophages. Moreover, lentiviral vectors have also proven to be effective in transducing brain, liver, muscle, and retina in vivo without toxicity or immune responses. The ibidi LifeAct-Lentiviral Vectors mediate efficient transduction, integration, and long-term expression of LifeAct into these cell types.

Fusion of a lentiviral vector with the cell membrane of mammalian cells (here rLV-LifeAct)

Method Comparison


Adenoviral Transduction

Lentiviral Transduction

Transient and stable protein expression

Transient protein expression

Stable protein expression

Easy-to-transfect cells (e.g., cell lines)

Difficult-to-transfect cells (e.g., primary cells)

Difficult-to-transfect cells (e.g., primary cells)

Variable transfection efficiency

Up to 100% transduction efficiency

Up to 100% transduction efficiency

Biosafety level 1

Biosafety level 2

Biosafety level 2

Transduction of Primary Endothelial Cells

The restricted uptake of viruses often lowers the success of gene or shRNA expression, respectively, and requires the application of higher viral titers. However, increasing the amount of viruses to boost expression often leads to toxic side effects in many cell types. ibidi’s ibiBoost Adenovirus Transduction Enhancer was developed to specifically enhance the efficiency of the virus transfer into difficult to transduce mammalian and rodent cells. By bridging the virus surface to the cell membrane, the virus uptake is significantly increased without toxicity.

Human umbilical vein endothelial cells (HUVEC) were transduced with the adenoviral vector rAV CMV-LifeAct-TagRFP and cultured under flow conditions (20 dyn / cm²) in a µ-Slide I 0.4 Luer over 9 days.