Cytoskeletal dynamics: - Actin dynamics in cellular processes - Live cell imagingand microscopy
Superb biocompatibility - non-toxic staining of living samples
Easy generation of stable cell lines
For in vitro and in vivo applications
1 x 107 TU / ml
TagGFP2 (Exmax / Emmax)
483 / 506 nm
TagRFP (Exmax / Emmax)
555 / 584 nm
NOTE: Please note that you will be working with a product containing infectious virus. Follow the recommended NIH guidelines for all materials containing BSL–2 organisms.
LifeAct, a 17-amino acid peptide, is derived from a protein from Saccharomyces cerevisiae. LifeAct stains filamentous actin (F-actin) structures in living or fixed eukaryotic cells and tissues. In contrast to GFP-actin and its alternatives, such as actin-binding proteins, LifeAct does not interfere with actin dynamics in vitro and in vivo, as shown in several cell types and model organisms.*
Lentiviral vectors derived from the human immunodeficiency virus (HIV-1) have become major tools for gene delivery into mammalian cells. The most advantageous feature of lentiviral vectors is the ability to mediate efficient transduction, integration, and long-term expression into dividing and non-dividing cells, both in vitro and in vivo.
The pseudotyped envelope with vesicular stomatitis virus envelope G (VSV-G) protein broadens the target cell range. Lentiviral vectors have now been shown to deliver genes to neurons, lymphocytes and macrophages. Previous retrovirus vectors could not be used for these cell types. Moreover, lentiviral vectors have proven to be effective in transducing brain, liver, muscle, and retina in vivo without any toxicity effects or immune responses.
The provided lentivirus is a VSV-G-pseudotyped pantropic virus, which is capable of infecting both dividing and non-dividing cells, while stably integrating into the cells’ genome.
After the transduction of cells with rLV-LifeAct, F-actin is visualized using the fluorescence markers TagGFP2 or TagRFP**. With this method, transduction efficiencies of up to 100% can be attained, even in difficult-to-transfect cell types like primary cells (e.g., neuronal cells). This virus can be used to generate stable cell lines, and stable clones can be selected by fluorescence sorting.
LifeAct plasmids may be used in many cell types when a transient or stable expression of LifeAct is needed. In this case, researchers can choose between two different promoters: CMV (cytomegalovirus) or CAG (modified chicken β-actin).
Adenoviral vectors are available for difficult-to-transfect cells, or when up to 100 % transgene expressing cells are needed. In this case, it is also possible to choose between the two promoters previously mentioned. These vectors are suitable for applications where only the transient expression of LifeAct-TagGFP2 or LifeAct-TagRFP is needed.
Method Comparison: Using LifeAct for F-Actin Visualization in Living Cells