Additional Immortalization Agents for Primary Cell Line Development

Additional immortalization agents are lentiviral and other recombinant constructs delivering genes beyond TERT and SV40 large T antigen used to immortalize primary cells, including HPV-16 E6/E7 (commonly used for epithelial and keratinocyte immortalization), Bmi1, cMyc, CDK4, and HOXA/HOXB family genes. They are supplied as lentiviral particles with antibiotic or fluorescent-antibiotic fusion selection markers. Academic researchers immortalizing epithelial cell types or exploring alternative immortalization mechanisms can use MBP's specialist team to confirm which agent fits a specific cell type.

Explore available immortalization agents or request a quotation by contacting customerservice@mbpinc.net. Our team can help identify the most suitable genetic construct and selection strategy for your specific cell type and experimental goal.

Additional Immortalization Agents

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What are additional immortalization agents?

Additional immortalization agents cover recombinant lentiviral constructs delivering genes other than TERT or SV40 large T antigen used to bypass replicative senescence in primary cells, most notably HPV-16 E6/E7 (which inactivate p53 and Rb similarly to SV40T but are derived from human papillomavirus), Bmi1 (a polycomb group protein that represses the p16/CDKN2A senescence pathway), cMyc, CDK4, and HOXA/HOXB family genes. Related entities include p16/CDKN2A pathway repression (Bmi1), keratinocyte and epithelial cell immortalization (HPV E6/E7), polycomb group signaling, and combination immortalization strategies.

What you will find:

Lenti-HPV-16 E6/E7 degrades Rb and p53 for enhanced cell immortalization.
Lenti-Myc T58A & Ras V12 enhance transformation and proliferation assays with high-titer mutant oncogenes
Lenti-p53 & Rb siRNA Virus extends cellular longevity and targets the knockdown of tumor suppressors
Lenti-HOX Series (RFP Tagged) for lineage-specific expansion and includes high-titer vectors like HOXB8, HOXA9, and HOXA10 vectors.
EBV Virus (Wild-Type) for immortalization workflows of B-lymphocytes
Lenti-CDK4 & Bmi1 Virus helps maintain proliferative capacity using cell mediators

How to choose additional immortalization agent products

HPV E6/E7 for epithelial and keratinocyte immortalization

HPV-16 E6 and E7 proteins inactivate p53 and Rb respectively, similar in net effect to SV40 large T antigen but derived from a different viral source; E6/E7-based immortalization is particularly well-documented for keratinocytes and other epithelial cell types, and is sometimes preferred over SV40T for these lineages based on published precedent.

Bmi1 for stem and progenitor cell types

Bmi1 represses the p16/CDKN2A senescence pathway through its role in the polycomb repressive complex, and has been used to extend the proliferative capacity of hematopoietic and neural stem/progenitor cells in research settings; confirm the specific cell type has documented precedent for Bmi1-based extension before relying on it as a primary immortalization strategy.

cMyc and CDK4 as proliferation-driving factors

cMyc and CDK4 overexpression can drive proliferation and contribute to immortalization, often used in combination rather than as standalone immortalization agents; review the specific combination strategy in the source literature before ordering individual components.

E6/E7 separate vs. fused constructs

HPV E6/E7 may be delivered as separate genes or as a fused open reading frame; confirm which configuration a published protocol specifies, since this can affect relative expression levels of the two proteins.

Specifications context

Additional immortalization agent lentiviruses are typically supplied at titers around 1x10^8 TU/mL, shipped on dry ice and stored at -80°C, with antibiotic or fluorescent-antibiotic fusion selection markers consistent with other immortalization lentivirus products. Pack sizes for academic labs commonly consist of small-volume aliquots sufficient for multiple transduction attempts, with constructs validated by sequencing and tested free of common contaminants.

This sub-category sits within MBP's cell immortalization catalog alongside TERT overexpression viruses and SV40T antigen overexpression viruses. Labs constructing custom combination vectors can also review MBP's cloning reagents. MBP's specialist team can help confirm which immortalization agent has documented precedent for a specific cell type before order placement.

Contact the expert team at MBP today to book high-quality immortalization vectors for your lab.

FAQ

HPV-16 E6 and E7 proteins inactivate the p53 and retinoblastoma (Rb) tumor suppressor pathways respectively, producing an effect similar to SV40 large T antigen but derived from human papillomavirus. HPV E6/E7-based immortalization is particularly well-documented for keratinocytes and other epithelial cell types.

Bmi1 is a polycomb group protein that represses the p16/CDKN2A senescence pathway, and has been used to extend the proliferative capacity of hematopoietic and neural stem/progenitor cells in research settings. Confirm the specific target cell type has documented precedent for Bmi1-based extension before relying on it as a primary immortalization strategy.

cMyc and CDK4 overexpression can drive proliferation and contribute to immortalization, but are more commonly used in combination with other factors such as TERT rather than as standalone immortalization agents. Reviewing the specific combination strategy described in the source literature for a target cell type is recommended before ordering individual components.

HPV E6/E7 may be delivered as separate genes or as a fused open reading frame, and the configuration can affect the relative expression levels of the two proteins. The published protocol being followed should specify which configuration was used, since this may affect immortalization efficiency for a specific epithelial cell type.

HPV E6/E7-based immortalization has substantial published precedent specifically for keratinocytes and other epithelial cell types, sometimes preferred over SV40 large T antigen for these lineages based on documented success rates. Both approaches inactivate similar tumor suppressor pathways but are derived from different viral sources.

Additional immortalization agent lentiviruses, including HPV E6/E7 and Bmi1, are commonly available with antibiotic resistance markers such as puromycin, hygromycin, and blasticidin, or fluorescent-antibiotic fusion markers. Confirm the selection marker does not conflict with any existing genetic modifications in the target cell line.

HPV E6/E7 expression is commonly confirmed by PCR detection of the integrated transgene, and functional confirmation may include assessment of p53 and Rb pathway status in the transduced cells compared to the original primary cells. Marker expression comparison to confirm retention of relevant epithelial identity markers is also commonly performed.

cMyc, CDK4, and other additional immortalization agents are frequently used in combination with hTERT in published immortalization strategies, particularly for cell types resistant to TERT alone. The specific combination and order of factor introduction should follow the source literature for the target cell type.

Additional immortalization agent lentiviruses, including HPV E6/E7, Bmi1, cMyc, and CDK4, are sold for research use only (RUO), and the resulting immortalized lines carry integrated viral genetic material generally unsuitable for direct therapeutic use without additional regulatory-grade development. Labs pursuing clinical-adjacent applications should plan for additional characterization beyond standard research immortalization.

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