Lentivirus Production

Lentivirus is a powerful gene delivery tool. Lentivirus can transduce most cells, including easy-to-transfect cells, hard-to-transfect cells, dividing, non-dividing and primary cells. The transduction efficiency can easily reach to 100% for most common cell lines.

What is lentivirus and how to make lentivirus? OriGene has a very informative lentivirus 101 ebook Which can quickly transform you into a lentivirus expert.

OriGene has genome-wide lentiviral vectors and shRNA lentiviral products. With the easy-to-use lentivirus production kits/reagents you can make high quality viral particles stress free.

Lentivirus Production

Lentivirus Titration

Lentivirus Concentration

Lentivirus Stabilization

Lentiviral transfer vectors (lenti vectors containing your gene) need essential viral genes to package into lentiviral particles. Lentiviral packaging genes are provided on separate plasmids – lentiviral packaging plasmids. OriGene’s Lenti-vpak packaging kit contains all necessary lentiviral production genes and is designed to package the third generation lentivectors into high titer viral particles, If you have your packaging plasmids, OriGene has a specialized lentivirus production transfection reagent, LentiTran, which is specifically formulated for high lentivirus production.

• Optimized for high packaging efficiency: Premix of the packaging plasmids at optimal ratio




• A potent transfection reagent included




• Simple to use: Transfect and harvest viral supernatant

HT1080 cells transduced with GFP lentiviral particles produced from Lenti-vpak packaging kit

For Regular Lentiviral Particle Packaging

Lentivirus produced will integrate into the host genome, enabling stable cell selection

TR30022 has been upgraded to TR30037.

For integration-deficient Lentiviral Particle (IDLV) Packaging

Does the viral titer need to be measured?

Virus titration will show how successful your lentivirus production is and will help to trouble shoot if viral transduction fails. Therefore virus titration is necessary. In addition, if your cells are hard to transduce and optimization is needed by using different MOI (Multiplicity Of Infection), the lentiviral titer needs to be measured.

What is MOI and how to calculate the amount of virus needed?

MOI is the number of viral particles per cell. (Recommended lentivirus MOI for common cell lines)

1. How to calculate the total number of viral particles for certain MOI?

If you want to use MOI of 10 and there are 10^4 cells in the well/dish, what is the total number of viral particles that are needed? The total viral particles is MOI x number of cells, 10 x 10^4 = 10^5 particles.

2. How to calculate the amount of virus needed per well?

If the lentiviral titer is 10^7 TU/mL, the amount of virus needed is 10^5/10^7 = 0.01mL

One-Wash™ Lentivirus Titer Kit

Features

• Simplified Workflow
  • No need for 37°C incubation
• Shortened protocol
  • One wash instead of 4 washes
  • Sample binding: 2 hrs vs overnight

If the lentiviral titer is low, the virus can be concentrated quickly with Lentiviral Concentrator, easily reaching to 10^9 TU/mL. If higher titer is needed or ultra pure virus is needed for in vivo animal studies, ultra centrifugation is needed. Custom lentivirus Production of flexible titers and volumes are provided from OriGene.

Lentiviral Concentrator Features

• Rescue low-titer viral particles, up to 100 fold
• Simple mix and centrifuge, no need for ultracentrifugation
• Easily scaled up for large volumes
• Protection from freeze-thaw after concentration
• Download Datasheet

lentiviral GFP particles (8 ml) were concentrated using TR30025, resuspended in 80ul of PBS.

GFP fluorescence image 72 hrs post transduction of HT1080 cells using lenti particles before and after concentration.

Lentiviral particles are enveloped viruses and tend to lose infectivity during storage, even at -80°C. OriGene developed a lentiviral storage buffer, Lentivirus Stabilizer, which can preserve the infectivity for at least one year at - 80°C.

Features

• Stabilizing lentivirus in storage
• Prolong storage, > 1 year
• Protect freeze and thaws: When used together with Lenti concentrator (TR30025)
• Download datasheet

Lentivirus Stabilizer Well Preserved the Infectivity of Lentivirus

Lenti-GFP particles (cat# PS100071V) freshly made (left panel). The viruses in culture media or Lenti Stabilizer were incubated at 37°C for 5 days. HEK293T cells were transduced with the above lentiviruses. Fluorescent images were taken 48 hrs post transduction.