Overexpression Cell Line Engineering Service
Custom Developed Protein Expression Cell Lines
You tell us the RefSeq or other target gene identifier and the cell line of interest.
Our partner Cellecta does the rest - they can make any cell into an overexpression cell using genomically integrating lentiviral cDNA vectors.
Lentiviral technology enables Cellecta to efficiently generate stable expression lines with almost any mammalian cell. A range of stable clones can easily be generated and then selected for moderate or high expressers, depending on the experimental requirements.
• Virtually any gene can be expressed in almost any mammalian cell line from nearly any cell type, including cells that are difficult to transfect and grow incl. primary cells
• Chromosomally integrated lentiviral cassette better mimics endogeneous gene expression than episomal constructs
• Able to generate overproducer cell lines with multiple integrated transgene constructs
• Express virtually any ORF with selection markers of your choice.
Lentiviral expression vectors are the most effective vehicles for delivering genetic material to almost any mammalian cell—including non-dividing cells and model organisms. A pseudoviral-packaged lentiviral cDNA construct can be highly efficiently transduced and integrated into virtually any mammalian cells—even the most difficult-to-transfect primary, stem, and differentiated cells.
Please check with us if your cell line of choice is in stock at Cellecta.
When it is not in stock, you can either provide it to us and we will ship it to Cellecta at a nominal shipping charge, or Cellecta can purchase it locally and we will charge you their purchasing costs.
You can order the engineering service in a modular way:
Stable Recombinant Protein Expression Cell Pool Construction
For the service we need the RefSeq # for the gene/protein of interest.
You can provide us with the gene, or we can select a clone from our various clone resources, or the gene is synthesized. (Cost depends - ask for a quotation).
The ORF (up to 3kb can be expressed efficiently with this system) can be cloned under control of CMV, EF1, or UbiC, depending on the expression level you want (e.g., high expression or moderate) and the types of cells (some promoters don’t work as well in some cell types). Constructs can also be designed to contain antibiotic selections (e.g. puro) and/or fluorescence markers.
For Cellecta vector information see link below.
After transduction of the lentiviral expression construct into your cell line, expression is selected and confirmed by either PCR or Western, as appropriate. After initial infection and integration with the expression construct, the result is a heterogeneous population–multiple cells with the genetic construct integrated at various locations in the genome. Different cells will express the protein at different levels so the culture is in fact a mix of different stable cells. Best candidates are expanded and a cell pool of 1 x 10e6 cells is obtained.
The whole process takes about 7-8 weeks.
Selection, Expansion, and Confirmation of Clonal Isolate from Cell Pool (Stable Recombinant Protein Expression Cell Line)
If the purpose is just to express a protein in a specific cell environment, a cell pool is sufficient. However, if the purpose is the investigate a certain cell process or response over several generations, it is preferable to obtain a homogeneous population, as a clonal population is usually very stable over time (unless the gene is particularly toxic or somehow unstable in the cells).
In this service module the cell pool is undergoing a selection process to establish stable monoclonal cultures each deriving from one sinle cell. These are amplified and expression is reconfirmed. A fully characterized stable clone is provided. You can let us know what kind of expression level is desired for your clone.
Depending on the growth rate of your cells this process takes about 6-8 weeks.
|Stable Recombinant Protein Expression Cell Pool Construction||CSCL-CD-X-GVO-CT||1 x 10^6 cells||please inquire €||DETAILS||Add to Cart|
|Selection, Expansion, and Confirmation of Clonal Isolate from Cell Pool (Stable Overexpression Cell Lines)||CSCL-XX-CP-GVO-CT||1 clone||please inquire €||DETAILS||Add to Cart|