Overexpression Stable Cell Line

VectorBuilder can engineer cell lines to stably over-express your gene of interest (GOI) to help you overcome variations associated with transient transfection. Gene overexpression cell lines are generated by lentiviral transduction followed by antibiotic selection. The GOI expression level is validated by RT-qPCR. Additionally, a series of standard QC assays such as sterility tests and mycoplasma detection are performed before releasing the final cell line products. Other validation assays such as Western blot and immunocytochemistry can be performed upon request.

Highlights

  • Permanent integration: lentiviral transduction allows for stable integration into the genome of host cells and all progeny.
  • Broad tropism: VSV-G pseudotyped lentivirus is able to bind to surface receptors on most mammalian cells, facilitating efficient transduction of your target cells.
  • Fast turnaround: engineered cells can be delivered in as fast as 9 weeks starting from vector design.

Service Details

Workflow for gene overexpression stable cell line engineering

Figure 1. Typical workflow of gene overexpression stable cell line production.

Price and turnaround
Service TypeDeliverablePrice (USD)*Turnaround
Gene overexpressionMixed pool (>106 cells/vial, 2 vials)From $2,9999-15 weeks
Two single clones (>106 cells/vial, 2 vials per clone)From $3,99914-20 weeks

* Additional charge will apply for extra single clones or vials.

QC assays
AssaysMethods
Overexpression validation (default)RT-qPCR
Expression test (add-on)WB, IF, FACS
Sterility (default)PCR for mycoplasma detection, bioburden test for sterility detection 

Downstream services

We can perform various phenotypic assessments and functional validation of your engineered cell lines, including assays for proliferation, apoptosis, migration, viability, cytotoxicity, etc. 

Case Studies

Validation of lentiviral-based gene overexpression

Figure 2. Validation of gene overexpression (OE) cell lines. (A) Transgene expression of ID8, CHO-K1, and HEK293 cells transduced with lentiviruses carrying MSLN, B7-H3, and CD19, respectively, analyzed by flow cytometry. (B) Protein expression level of GOI in Caco-2 cells by WB, and (C) luciferase overexpression in MCF7 cells by luciferase assay. (D) Luciferase-expressing Renca cell line-derived xenograft in mice were imaged to trace tumor migration in vivo.

FAQ

What are the advantages and disadvantages of overexpression vs. gene knockin?
ApproachAdvantagesDisadvantages
OverexpressionLentivirusTechnical simplicityRandom insertion
KnockinCRISPR (RNP electroporation)Targeted insertionMultiple components required
What are the advantages and disadvantages in using lentiviral transduction over plasmid transfection?
AdvantagesDisadvantages
Lentiviral transductionGenomic integration, permanent expression;
Ease of target cell transduction
Lower carrying capacity (6.4 kb)
Plasmid transfectionHigh carrying capacity (27 kb)Transient expression, plasmid will dilute after serial passages;
Requires electroporation or other transfection process
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