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Overexpression Cell Lines
Cyagen delivers targeted-gene-editing-optimized, GLP-compliant overexpression models validated by qPCR/Western blot, accelerating drug discovery from target screening (e.g., HER3, NEDD4) to IND-enabling studies. Backed by 2400+ peer-reviewed studies, with 8-12 week turnaround for academia and industry.
High-Expression Stability
Long-term expression using lentiviral integration ensures consistent results.
Customizable Promoters
Select from constitutive or tissue-specific promoters for targeted expression.
Lentiviral-Based Expression
Achieve stable gene overexpression across diverse mammalian cell lines.
Overview
Workflow
FAQs
Overview
Overexpression Cell Lines: Powering Breakthrough Research
Our overexpression cell lines deliver unparalleled genetic precision and experimental consistency. Leveraging cutting-edge lentiviral technology, we produce stable lines with up to 1,000-fold increased gene expression. Each model undergoes rigorous validation, ensuring reproducible results for your groundbreaking research in drug discovery and functional genomics.
Explore Ready-to-Use Mouse Models
Discover over 18,000 validated mouse strains—including knockout, conditional knockout, and humanized models—covering 20+ research areas such as oncology, neurology, and metabolism. All models are supported by detailed genotype data and guaranteed quality, helping you fast-track discovery with confidence.
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Workflow
Workflow and Delivery
Cyagen offers efficient overexpression cell line development services for gene function studies, drug screening, and disease model research. We provide both constitutive and inducible overexpression cell lines to meet diverse research needs. Each cell line is rigorously quality controlled and validated to ensure reliable experimental results.
Deliverables & Quality Control (QC)
Cell Type Example Cell Lines Deliverables Quality Control (QC)
Constitutive Overexpression HEK293, CHO, A549 - Stable cell pool or monoclonal cell lines (2 vials, 10⁶ cells/vial)
- Experimental report: Including vector report, experimental procedure, primer sequences, and transgene identification results.
- qPCR, Western blot (WB), Flow cytometry (FC), Immunofluorescence (IF)
- Functional assays: growth curves, cell cycle, proliferation, apoptosis
Inducible Overexpression HEK293, A549, U2OS
Note:
  • Cell Identification Report: Includes genotype analysis, primer sequences, vector report, and other essential details.
  • Experimental Report: Provides the detailed experimental procedure, gene editing results, cell line screening, and stability analysis.
  • Cell Line Delivery: Provides stable cell pools or monoclonal cell lines (2 vials, 10⁶ cells/vial).
Workflow & Timeline
Stage Description Timeframe
Cell Expansion & QC 1. Mycoplasma and sterility testing.
2. Cell proliferation assessment.
3. Primer design for sequencing the target region and confirmation of genotype.
1-2 weeks
Vector Construction 1. Gene synthesis
2. Subcloning of target gene into lentiviral expression vector
3. Preparation of expression plasmid
2-3 weeks
Lentivirus Packaging Co-transfection of target gene expression vector with helper plasmids into cells for lentivirus packaging. 2-3 weeks
Lentivirus Transduction & Drug Screening Transduce target cells with lentivirus vectors containing the target gene; select stable cells using appropriate antibiotics. 3-4 weeks
Expression Verification Verify the expression of the target gene or protein in stable cell lines and wild-type controls. 1 week
Note:Total Estimated Time: 9-13 weeks.
FAQs
Frequently Asked Questions (FAQs)
What solutions does Cyagen offer for these challenges?
Cyagen provides several solutions:
●Fusion tag technology: Proprietary hydrophilic fusion partners (e.g., YaiN/YbeL analogs) enhance solubility and purification efficiency for membrane proteins, achieving >80% success rates in functional reconstitution
●PTM-aware design: AI-driven sequence optimization identifies problematic PTM sites preemptively, enabling codon-optimized synthetic genes that bypass aggregation-prone regions while preserving functional domains
●Hybrid expression systems: Targeted-gene-editing-engineered HEK293 lines with inducible secretory pathway boosters enable authentic glycosylation and phosphorylation for therapeutic proteins
What are the main challenges when overexpressing membrane proteins or proteins with post-translational modifications?
The primary challenges include:
●Membrane protein insolubility: Hydrophobic regions often lead to aggregation, particularly in E. coli systems
●PTM incompatibility: Prokaryotic systems lack native eukaryotic post-translational modification machinery, risking misfolding and loss of functionality
●Cellular toxicity: Overburdened secretion pathways can impair cell viability during high-yield expression
What strategies do you employ to minimize cellular stress and toxicity when overexpressing proteins at high levels?
Cyagen implements several strategies:
●Inducible systems: Tetracycline- or ligand-regulated promoters allow controlled expression timing, reducing prolonged stress.
●Optimized delivery: Our 3rd-generation lentiviral systems achieve high transduction efficiency at lower viral titers, minimizing off-target effects.
●Promoter selection: Strong but regulated promoters like CAG enable high expression without requiring excessive plasmid DNA, lowering metabolic burden.
How do you optimize codon usage for the target gene to enhance expression levels in your chosen cell line?
We employ advanced multiparametric codon optimization algorithms that adjust codon bias to match the host cell’s translational machinery while stabilizing mRNA secondary structures. This approach, validated by industry-standard tools, increases protein yield without altering the amino acid sequence. For example, codon-optimized genes in HEK293 cells have demonstrated up to 86% improved expression in functional studies.
What are the most effective promoter systems for achieving high-level protein expression in mammalian cell lines?
Cyagen utilizes the CAG promoter (CMV enhancer fused to the chicken beta-actin promoter), which demonstrates superior performance in maintaining high-level, stable transgene expression compared to traditional promoters like CMV. This promoter drives stronger mRNA and protein yields in HEK293F cells, even under suboptimal conditions. For lentiviral-based systems, Cyagen’s targeted gene editing platform incorporates optimized promoters tailored to your specific cell line, ensuring broad tropism and minimal transcriptional silencing.
What Customers Say About Cyagen
Violet Shimmer
Stanford University
The service provided to us by Cyagen is now in press at Nature as an article.
Scarlett Rouge
Seattle Children’s Hospital
We are very pleased with the state-of-the-art professional transgenic services provided by Cyagen for our study published recently in Nature. We continue to use Cyagen’s transgenic services as it allows us to do better and more efficient research with transgenic mice.
Request a Custom Cell Line Consultation
Tell us about your cell line project needs. Our specialists are ready to support your research with tailored solutions.
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