Virus Packaging Process

If you need us to build lentiviral vector for your gene, please:

1. Provide a target gene sequence and the lentiviral vector information. We will synthesize the gene and subclone it for you.
2. If you have a target gene, please send us 1~5 µg of plasmid or bacterial culture for your gene.

We will subclone the gene of interest or shRNA into the lentiviral vector, prepare high quality plasmid DNA, transfect the packaging cell line, and harvest the lentivirus particle, followed by determining the concentration and titration. You will receive lentiviral vectors after titration.

If the lentiviral construct is ready, please provide us:

1. The construct map and sequence.
2. 1~5 µg of your lentiviral construct plasmid DNA or bacterial culture.

We will prepare endotoxin-free plasmid, transfect the packaging cell line for lentivirus production, and harvest the lentivirus particle, followed by determining the concentration and titration. You will receive lentiviral vectors after titration.

• Day1

  • Transfect 293F cells with combinations of packaging and lenti vectors

• Day4

  • Harvest conditioned medium

• Day5

  • TPurify or precipitate virus particles
  • Determine virus titers

Features

1. Chem-Lenti™ (in-house chemically defined media) is used to grow and transfect 293F cells. No animal component in crude virus harvest.
2. FusureTrans™ (innovative reagent for lentiviral packaging) for the transduction processes.
3. Consistent infectious units exceeding 1e7/ml virus in conditioned medium. We can further concentrate and purify the lentivirus.
4. Provide proprietary reagent to pull down viruses during the transduction process.
5. Employ in-house lentiviral vector for highly efficient packaging.

Innovations

Our team is dedicated to lentiviral vector development and virus packaging for genetic engineering. We now provide our superior lentiviral technologies for public use. Our PhD scientists have been dedicated to virus pseudotyping research for more than 15 years and developed unique technologies optimizing packaging efficiency, including in-house vectors, Chem-Lenti™, and FUSURE Trans™ Technologies.

Vector

We provide services from gene synthesis to lentivirus packaging services. Please note, use of our M1 Lenti Viral Vector will ensure consistent packaging results. Because packaging efficiency depends on the gene insert and type of lentiviral vector, we may not be able to assure consistent packaging using customer’s vector. For 1e7 titer range, the insert shall be less than 1.5 kb.

Chem-Lenti™ chemically defined media

Our in-house chemically defined media Chem-Lenti™ generates high titers of lentivirus.
Titers of the packaged virus are determined by serial dilutions and subsequent infection of 293F cells.
Titers are estimated by averaging cells infected when the infection ratios are in the 5-20% range. (See formula below for the 1:81 fold dilution)
Titer = 200,000 cells x 13.25% x 81 / 0.2 ml (virus dilution used) = 10.7 M/ml

Our lenti expression system provides rapid stable

We Produce Consistent Results

FuSure Reagent Pulls Down virus and Facilitates Transduction

Infection achieved by regular incubation for attached cells, or by 30 – 60 min spin at 300 g for suspension cells. No high speed spin is needed.