Our Custom Cell Line Development Services can generate stable cell lines for a wide variety of genes or siRNA sequences. From designing and synthesizing the requested cDNAs to editing your cell line, we offer end-to-end cell line development.
With our 10 years of experience developing stable cell lines, we have the latest technologies to guarantee the development of stable monoclonal cell lines, such as Cytena’s single-cell printer. But also guarantee the quality of the cell lines quantifying expression levels, making inducible the expression upon request, verifying the stability of the lines along the passes and supplying several clones with different levels of expression.
We can use any of our 60 parental cell lines, including the most requested CHO-K1, HEK293, SH-SY5Y, U2OS, HepG2 or Jurkat cell lines.
Stable Cell Line Development:
We offer stable Cell Line Development Service for a wide variety of genes including GPCRs, ion channels, proteases and kinases as well as various other enzymes, transmembrane proteins and adhesion molecules.
Expression of these genes could be a stable or inducible expression guaranteeing monoclonal cell lines using a single-cell printer from Cytena. Furthermore, at Innoprot we have developed a family of fluorescent reporter genes to measure Ca++, cAMP, DAG, RAS/RAF interaction, b-arrestin recruitment, Col1A1, etc.. which will allow you to perform directly your experiments on living cells.
Cell line development services include the next steps:
- Gene Synthesis
- Plasmid construction with the gene of interest
- Mammalian cell transfection with antibiotic selection
- Monoclonal Cells printing, screening and selection
- RT-PCR, IF study and Flow Cytometry Analysis
- Quantification of the number of receptors upon request
- Proof of cell stability during passages
- Selected clone expansion & Banking
- Mycoplasma, bacteria & Fungi screening
The time required to generate a new stable cell line is from 1 to 3 months. It depends on the cell line & the gene of interest.
Knockdown Cell Line Development:
We can generate knockdown stable cell lines with the type of silencing you’re interested in (inducible vs. constitutive). For this service, we use our functionally validated shRNA expression vectors.
Gene silencing by RNA Interference (RNAi) is a powerful research tool for studying gene function in mammalian cells. RNAi is a biological phenomenon by which double stranded RNA (dsRNA) specifically reduces gene expression of its corresponding gene.
We can develop them for a wide variety of genes including adhesion molecules, as well as various other enzymes. We introduce short interfering RNA (siRNA) and express gene products in the target cells, however the nucleic acids do not integrate into the host cell genome.
Knockdown Cell Line development service includes the next steps:
- Plasmid construction with the shRNA of interest
- Mammalian cell transfection and antibiotic selection
- Clone screening and selection
- Proof of cell viability, mycoplasma screening
- Clone expansion and banking
The time required to generate a new stable cell line is from 1 to 3 months. It depends on the cell line & the gene of interest.
Fluorescent Cell Line Development:
OurFluorescent Cell Line Development Service generates whole fluorescent stable cell lines by stably transfection with Fluorescent Proteins (FP). We can also generate stably expressing cells for a wide variety of genes or express vectors labelled with FPs. You only have to select the cell line and the FP and we will generate the recombinant cell line.
The following Fluorescent Proteins are available for your choice:
- Green Fluorescent Proteins: turboGFP & TagGFP2
- Red Fluorescent Proteins: TagRFP, turboFP602 & turboFP650
- Blue Fluorescent Proteins: TagBFP
- Yellow Fluorescent Proteins: turboYFP
If you are interested, we also develop fluorescent primary cells by transfecting FPs using lentiviral particles. Primary cells such as hepatic stellate cells, HUVEC, Neurons or Astrocytes are some examples of fluorescent primary cells we have already developed.