Cell Reprogramming & Transdifferentiation
Cellular reprogramming is the conversion of one cell type to another via the activation of gene networks that control a particular cell phenotype. This reprogramming technology represents a rapid way to generate target cells in both basic and clinical settings, which can be used for transplantation and studies of biology and diseases. Cellular reprogramming is typically achieved by overexpression of natural reprogramming factors (RFs) that control the gene networks corresponding to the desired cell phenotypes. The most prominent example of cellular reprogramming is the generation of induced pluripotent stem cells (iPSCs) from murine and human skin fibroblasts with four transcription factors (Oct4, Klf4, Sox2, c-Myc), which opened up the possibility of deriving different tissues by iPSC re-differentiation. Creative Biolabs has advanced platforms and cutting-edge technologies in the field of lentiviral vectors (LVs) design for cellular reprogramming. Our rapid and robust custom services of cellular reprogramming can streamline your research with an array of methods to meet your needs.
High Efficiency
Polycistronic vectors linked by 2A peptides ensure simultaneous expression of all reprogramming factors, increasing iPSC colony formation by 10-50 fold compared to separate vectors.
Excisable Safety
Integration of Cre-LoxP systems allows for the complete excision of viral cassettes after reprogramming, generating "minimal footprint" iPSCs suitable for disease modeling.
Direct Conversion
Custom vector design for transdifferentiation (e.g., Fibroblasts to Neurons) using lineage-specific factors like Ascl1, Brn2, Gata4, or Mef2c.
Reprogramming Vector Services
Polycistronic iPSC Vectors
Single-vector delivery of Oct4, Klf4, Sox2, and c-Myc (OKSM) for maximal stoichiometry.
We employ optimized constitutive polycistronic designs inspired by widely adopted 2A-linked reprogramming vector architectures, in which factors are separated by self-cleaving 2A peptides (P2A, T2A, E2A) to ensure equimolar expression.
Options to include Lin28 and Nanog (OKSMLN) for harder-to-reprogram cell types, or L-Myc variants for reduced tumorigenicity.
Validated on human fibroblasts, PBMCs, and urine-derived cells with >0.1% efficiency.
Minimal Footprint Excisable Systems
Generate preclinical-grade or disease-model-ready iPSCs without residual viral transgenes.
Reprogramming cassettes flanked by LoxP sites. Post-reprogramming, transient delivery of Cre recombinase excises the vector, leaving only a single LoxP site.
Includes antibiotic resistance or fluorescent markers (GFP/RFP) within the excisable region to easily identify successfully reprogrammed colonies before excision.
Direct Lineage Conversion
Transdifferentiation vectors to convert somatic cells directly into other lineages, bypassing the pluripotent state.
Vectors delivering BAM factors (Brn2, Ascl1, Myt1l) to convert fibroblasts into functional induced neurons.
Delivery of Gata4, Mef2c, and Tbx5 (GMT) for induced cardiomyocyte generation.
Reprogramming Enhancers
Accessory vectors to overcome epigenetic barriers.
Co-expression of epigenetic modifiers (e.g., Tet1/2, Kdm4) to facilitate chromatin opening and accelerate the reprogramming timeline.
Vectors expressing dominant-negative p53 or shRNA against p53/p21 to suppress senescence during the early phase of reprogramming.
Naïve State Induction
Tools for generating or maintaining cells in the "Naïve" ground state of pluripotency.
Vectors driven by specific enhancers (e.g., SRE, EOS) that are active only in the naïve state, coupled with antibiotic selection to purify ground-state colonies.
Technical Capabilities
Optimizing the reprogramming payload for maximum stability and minimal cytotoxicity.
Polycistronic Architecture
Balancing the expression levels of the four Yamanaka factors is critical for successful iPSC generation.
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2A Peptide Linkage We use optimized P2A, T2A, and E2A sequences to ensure high cleavage efficiency (>95%), preventing the formation of fusion proteins that could hinder nuclear translocation.
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Factor Ordering The order of genes (Oct4-Klf4-Sox2-cMyc) is empirically tested to account for the "position effect" in transcription, ensuring the correct stoichiometry for pluripotency induction.
Inducible Control
Precise temporal control over transgene expression allows researchers to turn off reprogramming factors once pluripotency is established.
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Tet-On Systems We incorporate 3rd generation Tet-On 3G promoters for tight, doxycycline-dependent regulation with minimal leakiness in the "Off" state.
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Safety Mechanisms Turning off c-Myc and other factors is crucial to prevent differentiation blockade and tumor formation in derived tissues.
Production & Quality
Reprogramming experiments often require high Multiplicity of Infection (MOI). We deliver concentrated virus to meet these demands.
| Parameter | Specification | Benefit for Reprogramming |
|---|---|---|
| Titer (Physical) | > 1 x 10^9 VP/mL | Allows for high MOI without adding excessive volume to culture media. |
| Purity | Ultra-purified (Chromatography) | Essential for sensitive primary cells (PBMCs, CD34+) to prevent toxicity. |
| Envelope | VSV-G (Standard) or Measles (Optional) | VSV-G provides broad tropism; other envelopes can target specific blood lineages. |
Designable Lentiviral Vector Types
We offer a flexible range of vector architectures optimized for reprogramming efficiency and safety. Choose the design that best fits your experimental timeline and downstream applications.
| Vector Type | Design Features | Application Context |
|---|---|---|
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Polycistronic Vectors (STEMCCA)
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Encodes all four reprogramming factors (Oct4, Klf4, Sox2, c-Myc) in a single cassette linked by self-cleaving 2A peptides. Driven by a strong constitutive promoter (e.g., EF1α). | Standard iPSC generation. Ensures equimolar expression of factors for maximal reprogramming efficiency in difficult-to-transduce cells. |
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Inducible Vectors (Tet-On 3G)
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Expression is driven by a promoter and controlled by Doxycycline (Dox). Includes a separate UbC-driven rtTA transactivator component for tight regulation. | Ideal for studying the kinetics of reprogramming, allowing researchers to turn off factors precisely after pluripotency is established. |
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Excisable Vectors (Cre-LoxP)
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The reprogramming cassette is flanked by LoxP sites. Post-reprogramming, transient delivery of Cre recombinase removes the viral DNA from the genome. | Generation of "minimal footprint" iPSCs for disease modeling or potential clinical applications where residual viral sequences are undesirable. |
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Reporter Vectors (GFP/RFP/Puro)
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Incorporates fluorescent proteins (GFP/RFP) or antibiotic resistance genes (Puro/Blast) linked to the reprogramming factors or driven by independent promoters. | Visual monitoring of transduction efficiency and easy selection of successfully transduced cells before colony formation begins. |
Need a specific combination not listed above?
Application Scenarios
From generating patient-specific iPSC banks to studying cell plasticity.
Patient-Specific iPSCs
Reprogramming somatic cells (fibroblasts, PBMCs) from patients with genetic disorders. The resulting iPSCs retain the patient's genotype, allowing for the creation of relevant disease models for drug screening.
Senescence Studies
Utilizing our specialized vectors containing Lin28 or shRNA-p53 to rejuvenate and reprogram aged or senescent cells, which are typically resistant to standard OKSM protocols.
Regenerative Medicine
Generation of minimal footprint iPSCs for potential autologous cell therapy. Our excisable systems ensure that no oncogenic viral sequences remain in the cells used for transplantation.
Transdifferentiation
Direct conversion of abundant somatic cells (e.g., skin fibroblasts) into scarce cell types (e.g., dopaminergic neurons or cardiomyocytes) for immediate use in assays, skipping the time-consuming iPSC stage.
Drug Screening
Leveraging iPSC-derived lineages (hepatocytes, cardiomyocytes) to construct physiologically relevant platforms for high-throughput compound screening and safety assessment.
Combining reprogramming with gene editing technologies to generate isogenic control lines or correct pathogenic mutations, facilitating precise functional genomic studies.
Service Workflow
Streamlined vector construction and packaging for iPSC generation.
Consultation & Strategy
We define the optimal factor combination (OKSM vs OKSMLN) and vector type (Constitutive vs Excisable) based on your target cell type and safety requirements.
Gene Synthesis & Cloning
Factors are synthesized and cloned into the lentiviral backbone, linked by 2A sequences. Restriction analysis and Sanger sequencing confirm plasmid integrity.
Virus Packaging
Vectors are packaged in HEK293T cells using a 3rd generation system. We use optimized transfection protocols to handle large polycistronic constructs.
Purification & Titration
Viral supernatant is concentrated via ultracentrifugation. Titers are verified by p24 ELISA and qPCR to ensure >10^8 - 10^9 TU/mL.
Delivery & Support
Vectors are shipped on dry ice. We provide detailed protocols for transduction and optional functional validation data (e.g., Alkaline Phosphatase staining of control cells).
What You Receive
Everything you need to start reprogramming immediately.
Transfer Plasmid
Complete sequence map and glycerol stock of your custom polycistronic reprogramming vector.
High-Titer Virus
Concentrated lentiviral particles (standard 100 µL - 1 mL aliquots) formulated in PBS/sucrose for maximum stability at -80°C.
QC Report
Certificate of Analysis showing titer (p24/qPCR), sterility (bacteria/fungi), and mycoplasma status.
Why Choose Lentivirus Today?
While non-integrating methods (Sendai, mRNA) exist, Excisable Lentiviral Vectors remain a widely adopted approach for balancing robust efficiency, broad cell tropism, and cost-effectiveness.
| Reprogramming Method | Efficiency | Footprint (Genomic) | Cost & Workload | Best Use Case |
|---|---|---|---|---|
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Lentivirus (Cre-LoxP)
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High |
Reduced footprint after excision. Single residual LoxP site remains. |
Cost-effective / Streamlined. Single infection event; stable reagents. |
Hard-to-reprogram cells (Blood, Aged cells); Large-scale banking. |
| Sendai Virus (SeV) | High |
Non-integrating RNA. Viral RNA clearance required. |
Higher material costs. Kits and licensing fees apply. |
Projects requiring non-integrating protocols. |
| Episomal Plasmids | Variable |
Non-integrating DNA. Dilutes out over cell divisions. |
Low reagent cost / Labor-intensive. Requires electroporation (variable viability). |
Budget-constrained projects; Easy-to-transfect cells. |
| mRNA / Protein | Low - Moderate |
Transient expression. No DNA intermediate. |
Demanding workflow. Requires daily/repeated transfections. |
Applications sensitive to any DNA introduction. |
Frequently Asked Questions
Plan Your Reprogramming Project
To ensure the best results, please consider the following when requesting a quote:
- Source Cells: Fibroblasts, PBMCs, or other?
- Method: Standard OKSM or Enhanced (Lin28/Nanog)?
- Safety: Do you need excisable (Cre-LoxP) vectors?
- Scale: Pilot study or large-scale banking?
- Control: Constitutive or Dox-inducible?
Get a Custom Vector Quote
Our vector design experts are ready to assist you in selecting the most efficient reprogramming tools.
Start Your Project Today
Tell us about your project, and our experts will get back to you with a customized quote and proposal.
