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Home | Viral Packaging | Trans-Lentiviral™ Packaging Sy | Biosafety Features

Biosafety Features
the safest lentiviral system for expression

The Trans-Lentiviral Packaging System is based on new generation lentiviral vectors developed by Kappes et al (1,2,3,4). This lentiviral system includes a significant number of safety features designed to enhance its biosafety and to minimize its relation to the wild-type, human HIV-1 virus. Figure 1 illustrates the absence of functional gag-pol recombinants in lentiviral stocks generated from the Trans-Lentiviral Packaging System.

Trans-Lentiviral Packaging System includes the following key safety features:

The expression vectors contain a deletion in the 3' LTR that does not affect generation of the viral genome in the producer cell line, but results in “self-inactivation” of the lentivirus after transduction of the target cell (5).
The number of genes from HIV-1 that are used in the system has been reduced (i.e. gag, pol, rev, tat and vpr).
None of the structural genes are actually present in the packaged viral genome, therefore no new replication-competent virus can be produced.
The VSV-G gene from Vesicular Stomatitis Virus is used to pseudotype the vector particles (Yee et al., 1994). The HIV-1 envelope has been completely removed from the vector.
Genes encoding the structural and other components required for packaging the viral genome are separated onto five plasmids minimizing the threat of recombinant replication competent virus production.
Although the four packaging plasmids allow expression in trans of proteins required to produce viral progeny (e.g. gag, pol, rev, env) in the TLA-HEK293T producer cell line, none of them contain LTRs or the Ψ packaging sequence. This means that none of the HIV-1 structural genes are actually present in the packaged viral genome, and thus, are never expressed in the transduced target cell. No new replication-competent virus can be produced.
The reverse transcriptase (RT) and integrase (IN) proteins are provided in trans producing a class of vectors that contain split gag-pol components on separate vectors

Safety Classification

This product is classified as Biosafety Level 2. For more information regarding viral agents and Biosafety Level 2 laboratory guidelines and precautions, please refer to the links below:

NIH Agent Summary Statement
NIH Biosafety Level 2 Description

Shipping Information:

References:

1. Kappes J.C., Wu X., Wakefield J.K. Production of trans-lentiviral vector with predictable safety. Methods Mol Med. 2003; 76:449-65.

2. Kappes J.C., Wu X. Safety considerations in vector development. Somat Cell Mol Genet. 2001; 26(1-6):147-58.

3. Wu X., Wakefield J.K., Liu H., Kappes J.C. Analysis of lenti- and trans-lentiviral vector genetic recombination. Dev Biol (Basel). 2001; 106:237-48; discussion 249, 253-63. PMID: 11761237

4. Wu X., Wakefield J.K., Liu H., Xiao H., Kralovics R., Prchal J.T., Kappes J.C. Development of a novel trans-lentiviral vector that affords predictable safety. Mol Ther. 2000; 2(1):47-55. PMID: 10899827.
5. Zufferey R., Dull T., Mandel R.J., et al. Self-inactivating lentivirus vector for safe and efficient in vivo gene therapy. J Virol 1998; 72:9873-9880.

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