Figure 1: Packaging with the Trans-Lentiviral System
NEWEST Generation Packaging System
The Trans-Lentiviral Packaging System is the most effective strategy for efficiently expressing your gene or shRNA of interest. Co-transfection of the plasmids into the packaging cell line, TLA-HEK293T™, allows efficient production of lentiviral supernatant which can then be transduced and expressed in a wider range of cell types than traditional retroviral systems (Figure 1).
Highly Efficient Expression
- CONSISTENT TITERS: 1 - 5 x 106 transducing units (TU) per ml (unconcentrated) are commonly achieved.
- BROAD TROPISM: Effectively transduce most mammalian cell lines including primary and non-dividing cells.
Multi-Generational Flexibility
The only commercially available packaging system that allows you to effectively package both second and third generation lentiviral vectors.
Unsurpassed Biosafety
Open Bisoystems engineered this new generation Trans-Lentiviral Packaging System to be effective and safe. The packaging system includes a number of features designed to enhance its biosafety and to prevent the generation of replication-competent viral particles. Additional information on the packaging mix plasmids and their safety features can be found at the Trans-Lentiviral Biosafety page.
System Components
The packaging system contains all of the necessary components for up to 10 packaging events and can be ordered with (system 2) or without (system 1) the packaging cell line, TLA-HEK293T. Or choose from two bulk packaging systems that contain a larger volume of the packaging mix and transfection reagent. The two bulk packaging systems have a sufficient quantity to perform 50 or 100 packaging events. The complete system includes the following components:
|
Cell Line* | TLA-HEK293T™ | Allows production of the lentivirus following co-transfection of the transfer plasmid and the packaging plasmids in the packaging mix |
Packaging Mix | Optimized mixture of the five packaging plasmids: pTLA1-Pak, pTLA1-Enz, pTLA1-Env, pTLA1-Rev and pTLA1-TOFF | Supply the helper functions as well as structural and enzymatic proteins in trans required to produce the lentivirus |
Transfection Reagent* | | Developed and optimized for transfection of plasmid DNA into the nucleus of cultured eukaryotic cells |
Vectors* (control and cloning) | pGIPZ™ Non-Silencing Control Vector (shRNA) | Allows packaging of the expression construct into virions |
*For components that can be ordered separately
Gene Content Compatible with the Trans-Lentiviral Packaging System
Open Biosystems offers several gene content options for use in gene knockdown or gene expression using the Trans-Lentiviral Packaging System. We offer both shRNA and shRNAmir constructs for gene knockdown. These genome-wide libraries are available for both human and mouse, and incorporate several features aimed at increasing the efficiency and specificity. Human LentiORF Clones are also available for gene expression assays. The LentiORFs were specifically designed for transient and stable transfection and for lentiviral production.
Which kit to choose?
The Trans-Lentiviral pGIPZ Packaging System is designed to create replication incompetent virus for use in shRNA experiments.
The Trans-Lentiviral pLEX Packaging System is designed to create replication incompetent virus for use in overexpression experiments.
shRNA and shRNAmir products are covered by a Limited Use License.
These components have been validated and packaged along with detailed and simplified protocols. All plasmids are shipped on wet ice and stored at -20°C. Transfection reagents are shipped on wet ice and stored at 4°C. For kits containing the packaging cell line, TLA-HEK293T, an additional dry ice cooler will be included in the shipment. Upon receipt, the cell line must be stored at -80°C.
1. Kappes & Wu, Somatic Cell and Molecular Genetics, Vol. 26, Nos. 1/6, November 2001.
2. Kappes JC, Wu X, Wakefield JK. Production of trans-lentiviral vector with predictable safety. Methods Mol Med. 2003; 76:449-65.
3. Kappes JC, Wu X. Safety considerations in vector development. Somat Cell Mol Genet. 2001 Nov;26(1-6):147-58
4. Liu H, Wu X, Xiao H, et al. Incorporation of functional human immunodeficiency virus type 1 integrase into virions independent of the Gag-Pol precursor protein. J Virol 1997; 71:7701-7710.
