Generate the constitutive or inducible expression stable cells in your desired host cell line for your desired target
Mammalian stable cell line creation:
GenTarget’s stable cell line service provides a very cost-effective way for you to generate a cell line. Using our proprietary lentiviral vectors, we will subclone your target gene into our lentivector, generate the lentivirus particles, sequentially infect the cell line of your interest, and select the cell clones via Cell sorting and antibiotic killing.
For an inducible expression cell line, we will first transduce the cells with TetR repressor lentivirus, to make a TetR stable cell line, then transduce with the target expression lentivirus, to generate the tetracycline-inducible stable cell line.
Our proprietary suCMV promoter has the strongest expression strength in the marker in most cell types, and will provide extremely high levels of expression for your target. Our modified EF1a promoter will express the high level of target in some cell types that has weak CMV promoter strength. Our unique technology ensures a fast turnaround time. We will work with you to ensure that the cell line meets your requirements so that you will get the best quality and the best price.
Service Procedures of GenTarget Stable Cell line :
- Construct a lentiviral vector containing your gene and desired selection marker
- Generate lentivirus
- Transduce the cell line your desired
- Select the stably transduced
- Validate integration of the gene via genomic PCR
- Validate the highest-expressing clone by Western Blot (if applicable)
- Deliver two cryogenic vials of stable cells along with a final report
What are the advantages of using lentivirus to generate stable cells?
To make stable a cell line, the target has to be integrated into the host cells’ genome for stable, constitutive expression. Random integration (such as by transfection) demonstrates widely varying levels of expression depending on the integration site. Additionally, random integration often results in independent integration of the target and the selection marker, necessitating large-scale screening of select positive clones. Typically less than ten percent of antibiotic-resistant clones express the transgene. In contrast, lentivirus transduction tends to integrate at high transcription level sites (hot-spots) with a full virus genome. Engineered lentiviral transfer vector that is embedded with a matrix-attachment region (MARs) sequence may provide position-independent transgene expression. Compared to conventional stable cell line construction, transduction with lentivirus produces a much higher positive clone rate and the target always co-exists with the selection marker, thereby substantially reducing cost, labor, and time spent selecting stable high expression clones.
Why You Should Use GenTarget’s Stable Cell Line Service?
- Engineered in-house lentiviral vectors provide high-efficiency gene integration with extremely low false positives
- Safe, self-inactivating, high-titer lentiviral particles can deliver your gene into a wide range of cell lines including non-dividing, primary, or stem cells
- Target protein expression can be tracked and monitored by fluorescence imaging, allowing sorting or selecting of integrated cells (the fluorescent protein is not fusioned with your target).
- Our experts have years of experience in lentiviral cloning and expression
- Fast turnaround time
- The best price
How to order?
Please check our catalog products for pre-made Stable Cell Lines!