What is stable cell line generation (or stable cell line development)? Transferring genes (ORFs) into target cells to observe phenotypic changes is one of the common methods for gene function research. The basic principle of establishing a lentivirus stable cell line is to clone exogenous DNA to a vector with a certain resistance gene, the vector is then transfected/transduced into the host cell and target ORF is integrated into the host chromosome, and the resistance markers contained in the vector are used for positive cell screening.

▍  Lentivirus-mediated Stable Expression

The most common method for stable cell line construction is lentivirus transduction (ie. lentivirus stable cell line). This method overcomes the disadvantages of the traditional method of plasmid selection and cloning, and can obtain stable cell lines efficiently in a short time. The general resistance markers include neomycin, hygromycin and purinomycin. The cell lines that can stably express the target protein are screened by corresponding antibiotic.

With many years of experience in cell biology, Vitro Biotech provides gene overexpression stable cell line generation service. Multi-clonal stable cell pool or monoclonal stable cell line are all feasible! Feel free to contact us to get a strategy for your overexpression project.

Stable cell line generation workflow

▍  Transposon-mediated Stable Expression

The target CDS size exceeds 3kb? Try the transposon-mediated PiggyBac method. Under the catalysis of transposase, the target gene is cut off from the vector and inserted into the position with the certain sequence in the genome to achieve stable expression. This method has the advantages of low cost, high efficiency and long-term protein expression.


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