This short video is about lentivirus productions.
Why use lenti as a gene delivery tool? For many cells, such as primary cells or stem cells, regular chemical-based transfection reagents fail to deliver genes into those cells. Lentivirus, however, can efficiently deliver genes into them.
OriGene is a Lenti leader and offers genome-wide lenti products for human, mouse and rat. We also offer a series of lenti kits to help lenti production in your own lab.
Two Lenti product lines are available: Lenti-ORF for gene overexpression, or Lenti shRNA for gene knockdown.
For each Lentiviral products, there are two options. 1: lentiviral plasmids; 2: lentiviral particles.
For option 2, ready-to-use particles will be delivered and you can skip all the steps below.
For option 1, then you would need a few more steps viral particle production: packaging, titration, concentration and storage.
To package lentivirus, co-transfect a lenti vector, either lenti-ORF or Lenti-shRNA with a set of packaging plasmids into HEK293T cells. Lentivirus can then be harvested from the supernatant.
OriGene’s lenti packaging kit is the perfect tool for this job. It contains one vial of lyophilized lenti packaging plasmids, mixed at an optimal ratio. A vial of Turbofectin, a highly effective transfection reagent is also included.
After virus production, lenti titer kit, p24 ELISA can be used to measure the titer. P24 is the viral capsid protein, and there are around two thousand p24 molecules per virus. Therefore, p24 protein level can be converted into the amount of viral particles.
This diagram shows how the P24 ELISA works. P24 capture antibody is coated on a plate. P24 from the lentivirus will bind to the capture antibody, and then conjugated detection antibody is added. And using the microplate reader, the amount of P24 will be measured.
OriGene’s One-wash Lenti titer kit is the kit of choice, as its simplified protocol eliminates the 3 out of the 4 wash steps in conventional titering kits. In addition, it provides the benefits of room temperature process and shortened the time requirement to 3 hrs.
Now we come to the concentration step. Lentiviruses are more stable when concentrated and some applications require virus of higher titer. With OriGene’s Lenti concentrator, this can be done easily. Just mix viral supernatant and spin at low speeds. No need for an ultracentrifuge.
Lentivirus tends to lose infectivity in storage, even when stored at -80C. OriGene has developed a Lenti Stabilizer to tackle this problem. When the concentrated virus are stored in stabilizer solution, it extends the infectivity to 12 month from the regularly less than 6 months and protects the virus from multiple freeze-and-thaw. All of OriGene’s lenti particles are stored in this stabilizer solution.
In OriGene we create tools to facilitate research and to make your Lenti experience simple and enjoyable. For more details, please visit www.origene.com/lentiviral