LentiPlus Pseudovirus Assembly and Infection Kit

Product Description

LentiPlus Pseudovirus Assembly and Infection Kit

For Highly Efficiency Lentiviral Particle Assembly & Infection

The LentiPlus Pseudovirus Assembly and Infection Kit provide a convenient and efficient tool for producing lentiviral particles to deliver and express your gene of interest in dividing, non-dividing, or primary cells. The multi-vector pre-mixed Packaging Mix is easy to use and designed to express viral structural proteins and an envelope protein of your choice at ratios optimal for lentiviral assembly and production. Choose from over 14 viral envelopes to further optimize transduction to your target cell type. In addition, the Kit also provides a lentiviral transduction enhancer, Infectin^TM, a proprietary technology developed at Virongy, to significantly increase the productive viral transduction rate (up to 50-fold). Infectin^TM paired with the VSV-G (vesicular stomatitis virus glycoprotein) envelope protein for transduction efficiencies as high as 99% in certain cell types. The use of Infectin^TM eliminates the need to concentrate viral particles, and greatly facilitates biological and biochemical characterization of lentiviral transduced cells.

Highlights

• Pre-opitimized vector mix, ready to use, and easy steps
• Higher yield
• Infectin^TM technology that enhances transduction efficiency up to 30 fold
• No need for viral particle concentration
• High transduction efficiency for both cell lines and primary cells.

Kit Components of the LentiPlus Pseudovirus Assembly and Infection Kit

(with Tranfectin and Lenti-Infectin^TM reagents)

Example Assembly Protocol:

Lentiviral particle assembly using Lenti2go Lentiviral Kit (10 cm dish)

1. One day (18-24 hours) prior to co-transfection, plate 4 x 10⁶ HEK293T cells in a 10 cm dish, and grow at 37°C to reach about 80% confluence.
2. Remove medium from the dish and rinse with serum free DMEM medium, then add 9 ml of warm, serum-free medium.
3. Prepare transfection solution: Set up two tubes (A) and (B). (A) is for DNA dilution, and (B) is for dilution of Transfectin.
4. In (A), add 10 μg of LentiPlus Master Mix, add 10 µg of Lenti-viral vector of your choice, and then add serum-free DMEM medium to make a final volume of 500 μl, Mix well.
5. In (B), add 45 μl of Transfectin, and then add 455 μl if serum-free DMEM medium to make a final volume of 500 μl. Mix well.
6. Combine (A) and (B) into a total volume of 1 ml. Mix DNA and Transfectin by votexing, and incubate at room temperature for 15 min.
7. Add the 1 ml (A) + (B) mixture to the dish drop-wise. Mix gently by slowly rotating the dish. Incubate at 37°C for 6 hours.
8. Carefully remove the supernatant with a pipette (not to remove cells). Add 10 ml of warm DMEM with serum. Culture cells for 48 hours
9. Harvesting virus at 48 hours. Filter through a 0.45 μm filter to remove cellular debris. Aliquot and store the virus at -80°C.