LentiArt™ Virus Packaging Kit (CART-027CL)

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Product Name
LentiArt™ Virus Packaging Kit

Cat. No.
CART-027CL

General Description
Lentivirus vector based on the human immunodeficiency virus-1 (HIV-1) has become a promising vector for gene transfer studies. The advantageous feature of lentivirus vector is the ability of gene transfer and integration into dividing and non-dividing cells. The pseudotyped envelope with vesicular stomatitis virus envelope G (VSV-G) protein broadens the target cell range. Lentiviral vectors have been shown to deliver genes to neurons, lymphocytes and macrophages, cell types that previous retrovirus vectors could not be used. Lentiviral vectors have also proven to be effective in transducing brain, liver, muscle, and retina in vivo without toxicity or immune responses. Recently, the lentivirus system is widely used to integrate siRNA efficiently in a wide variety of cell lines and primary cells both in vitro and in vivo.
Lentivirus particles are produced from 293T cells through transient transfection of plasmids that encode for the components of the virion. Due to safety concerns regarding the infectious nature of HIV-1, recent lentiviral packaging systems have separated the viral components into 3 or 4 plasmids. However, these systems still present a small chance of generating replication-competent lentivirus upon recombination. In addition, most commercial lentiviral packaging systems provide plasmids containing the viral structure proteins in a premixed formulation, making it nearly impossible to optimize the ratio of the various plasmids for your particular experiment and host cell.
LentiArt™ Virus Packaging Kit provides a much safer method to package lentivirus, while still providing high viral titers. In addition, each plasmid is provided separately rather than in a packaging mixture. This allows you the flexibility to amplify individual plasmids and optimize the ratio of plasmids for your experiment.

Components
LentiArt™ pHelp1, 1ug/uL; 10μL; Store at -20°C;
LentiArt™ pHelp2, 1ug/uL; 10μL; Store at -20°C;
LentiArt™ pHelp3, 1ug/uL; 10μL; Store at -20°C;

LentiArt™ pHelp1: (9118 bp, Ampicillin-resistant). Pst I Digestion: 927 bp + 1424 bp + 6767 bp.
LentiArt™ pHelp2: (5822 bp, Ampicillin-resistant). EcoRI Didestion: 1668 bp + 4154bp.
LentiArt™ pHelp3: (4180 bp, Ampicillin-resistant). EcoRI Digestion: 300 bp + 3880 bp

Storage Conditions
Upon receipt, store all other kit components at -20°C until their expiration dates.

Procedure
1. One day before transfection, plate sufficient 293T cells to achieve 70-80% confluence on the day of transfection.
2. Transfect cells by Calcium Phosphate or other transfection reagents.

Note: We suggest transfecting cells with FuGENE® Transfection Reagent (Roche Applied Science) or Lipofectamine™ Plus (Invitrogen). We recommend the ratio of vectors at 3:1:1:1 (transfer vector: pHelp1: pHelp2: pHelp3).

3. Harvest lentiviral supernatant 36-72 hours after transfection. Supernatant can be harvested 2 or 3 times, every 12 hours. Keep it at 4°C over the collecting period.
4. Pool the collected supernatants, centrifuge 5 minutes at 1500 rpm to remove cell debris and filtrate on 0.22 µm.
5. Supernatants can be used directly or purified/concentrated if needed. For long term storage, store supernatant at -80°C in aliquots.

Product Use Statement
Remember that you will be working with samples containing infectious virus. Follow the recommended NIH guidelines for all materials containing BSL-2 organisms. The LentiArt™ Virus Packaging Kit is designed to minimize the chance of generating replication competent lentivirus, but precautions should still be taken to avoid direct contact with viral supernatants.

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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

LentiArt™ Virus Packaging Kit (CART-027CL)

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