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Cell Line Generation / Transfection

Generation of Stable Cell Line in 28 Days

Stable cell lines over-expressing a protein of interest (or shRNA construct targeting a gene of interest) are important research tools, particularly for the production of recombinant antibodies and other proteins, the study of gene functions, drug screening, and other applications. In addition, utilizing a stable mammalian cell line can make products more acceptable for clinical use. Stable cell lines will reliably grow for extended periods of time and can continue to express a transgene at a consistent rate. However, production of stable cancer cell lines and primary cells can be very expensive, labor-intensive, and time-consuming.
Sunomix Biosciences offers several stable cell line generation services: protein overexpressing cell lines, RNAi knockdown cell lines, and reporter stable cell lines (luciferase, GFP, RFP, YFP). Company scientists provide expert services with years of cell culture experience with more than 150 cancer cell lines (available in house). All clonal stable cell lines are developed based on individual client specifications, using highly efficient gene delivery technologies to produce stably expressing clonal cell lines in just 28 days. We can produce multiple clonal cell lines with various expression levels (low / medium / high expression).
Typically, antibiotic resistance or fluorescent reporter gene markers are incorporated into the plasmid DNA construct to facilitate selection process. These selection markers can be co-expressed on the same vector or independently expressed on two separate vectors. The selection process facilitates the selection of the most efficient expressers or silencers of the gene of interest.
Standard services include transfection of plasmid DNA (10 – 20 µg), drug selection of clonal cells, colony pick up, generation of a stable line, expression and functional screening (at least 10 passages), and final validation of construct expression by qRT-PCR and/or Western blot analysis.
The Western blot analytical technique uses antibodies specific to the target protein in order to quantify the expression of the protein in a sample. The genomic integration of the target gene and associated mRNA expression increase can be verified using quantitative real-time RT-PCR technique. The utilization of both techniques validate that the target gene is both integrated into the genome and functionally expressing.

To find out more details about this custom service, Please request a quote today.